Hepatoma-Derived Growth Factor: Its Possible Involvement in the Progression of Hepatocellular Carcinoma

The development of hepatocellular carcinoma (HCC) is an important complication of viral infection induced by hepatitis virus C, and our major research theme is to identify a new growth factor related to the progression of HCC. HDGF (hepatoma-derived growth factor) is a novel growth factor that belongs to a new gene family. HDGF was initially purified from the conditioned medium of a hepatoma cell line. HDGF promotes cellular proliferation as a DNA binding nuclear factor and a secreted protein acting via a receptor-mediated pathway. HDGF is a unique multi-functional protein that can function as a growth factor, angiogenic factor and anti-apoptotic factor and it participates in the development and progression of various malignant diseases. The expression level of HDGF may be an independent prognostic factor for predicting the disease-free and overall survival in patients with various malignancies, including HCC. Furthermore, the overexpression of HDGF promotes the proliferation of HCC cells, while a reduction in the HDGF expression inhibits the proliferation of HCC cells. This article provides an overview of the characteristics of HDGF and describes the potential role of HDGF as a growth-promoting factor for HCC.     

Effect of native microflora on the growth kinetics of salmonella enteritidis strain 04-137 in raw ground chicken

Effects of native microflora (NM) on growth kinetics of Salmonella Enteritidis strain 04-137 were studied in raw ground chicken. First, samples of ground chicken with high and low levels of NM (10(7.1) and 10(4.9) CFU/g, respectively) were spiked with Salmonella at doses ranging from 10(1) to 10(4) CFU/g. The growth kinetics, including the rate constant of growth, r, and the lag period, were similar, but the maximum cell level, N(max), was higher at higher initial Salmonella doses for both NM levels. Second, samples of ground chicken with high and low NM levels (10(6.8) and 10(4.7) CFU/g, respectively) were spiked with Salmonella and then stored at various constant temperatures ranging from 8 to 32°C. Both N(max) and r for Salmonella were higher at higher temperatures for both NM levels. Although r for total bacteria, which consisted of NM and Salmonella, was also higher at higher temperatures, N(max) was constant at all temperatures for both NM levels. Further, Salmonella growth was compared among samples of ground chicken with high and low NM levels and samples of sterilized chicken. Salmonella growth, characterized by both N(max) and r, was highest in sterilized chicken, followed by chicken with the low NM level. Our growth model successfully described and analyzed the growth of Salmonella and total bacteria in chicken at constant temperatures; using the data obtained, the model also successfully predicted the growth of Salmonella and total bacteria in chicken stored at dynamic temperatures. Our study clarified the effects that different doses of NM in ground chicken had on the growth kinetics of the Salmonella strain and demonstrated the usability of the growth model for foods with NM.     

Exendin-4 Promotes Schwann Cell Survival/Migration and Myelination In Vitro

Besides its insulinotropic actions on pancreatic β cells, neuroprotective activities of glucagon-like peptide-1 (GLP-1) have attracted attention. The efficacy of a GLP-1 receptor (GLP-1R) agonist exendin-4 (Ex-4) for functional repair after sciatic nerve injury and amelioration of diabetic peripheral neuropathy (DPN) has been reported; however, the underlying mechanisms remain unclear. In this study, the bioactivities of Ex-4 on immortalized adult rat Schwann cells IFRS1 and adult rat dorsal root ganglion (DRG) neuron–IFRS1 co-culture system were investigated. Localization of GLP-1R in both DRG neurons and IFRS1 cells were confirmed using knockout-validated monoclonal Mab7F38 antibody. Treatment with 100 nM Ex-4 significantly enhanced survival/proliferation and migration of IFRS1 cells, as well as stimulated the movement of IFRS1 cells toward neurites emerging from DRG neuron cell bodies in the co-culture with the upregulation of myelin protein 22 and myelin protein zero. Because Ex-4 induced phosphorylation of serine/threonine-specific protein kinase AKT in these cells and its effects on DRG neurons and IFRS1 cells were attenuated by phosphatidyl inositol-3′-phosphate-kinase (PI3K) inhibitor LY294002, Ex-4 might act on both cells to activate PI3K/AKT signaling pathway, thereby promoting myelination in the co-culture. These findings imply the potential efficacy of Ex-4 toward DPN and other peripheral nerve lesions.     

Characterization of the precursor of prostate-specific antigen. Activation by trypsin and by human glandular kallikrein

The precursor or zymogen form of prostate-specific antigen (pro-PSA) is composed of 244 amino acid residues including an amino-terminal propiece of 7 amino acids. Recombinant pro-PSA was expressed in Escherichia coli, isolated from inclusion bodies, refolded, and purified. The zymogen was readily activated by trypsin at a weight ratio of 50:1 to generate PSA, a serine protease that cleaves the chromogenic chymotrypsin substrate 3-carbomethoxypropionyl-L-arginyl-L-prolyl-L-tyrosine-p-nitroanili ne- HCl (S-2586). In this activation, the amino-terminal propiece Ala-Pro-Leu-Ile-Leu-Ser-Arg was released by cleavage at the Arg-Ile peptide bond. The recombinant pro-PSA was also activated by recombinant human glandular kallikrein, another prostate-specific serine protease, as well as by a partially purified protease(s) from seminal plasma. The recombinant PSA was inhibited by alpha1-antichymotrypsin, forming an equimolar complex with a molecular mass of approximately 100 kDa. The recombinant PSA failed to activate single chain urokinase-type plasminogen activator, in contrast to the recombinant hK2, which readily activated single chain urokinase-type plasminogen activator. These results indicate that pro-PSA is converted to an active serine protease by minor proteolysis analogous to the activation of many of the proteases present in blood, pancreas, and other tissues. Furthermore, PSA is probably generated by a cascade system involving a series of precursor proteins. These proteins may interact in a stepwise manner similar to the generation of plasmin during fibrinolysis or thrombin during blood coagulation.     

Prognostic criteria in patients with renal pelvic and ureteral cancers--clinical value of volume weighted mean nuclear volume

A retrospective, prognostic study was performed on 22 patients with transitional cell carcinoma of the upper urinary tract seen between Dec. 1982 and Jun. 1995. Unbiased estimates of volume weighted mean nuclear volume (MNV) were compared with age at the time of diagnosis, muscle invasion, and histological grading according to World Health Organization (WHO) classification on the prognostic value. Estimates of MNV were significantly correlated (p = 0.0135) only with disease-specific survival of transitional cell carcinoma of the upper urinary tract, contrary to age at the time of diagnosis (p = 0.4865), and muscle invasiveness (p = 0.1756). The analysis was not conclusive with regard to the WHO classification as the prognostic factor in this study. Estimates of MNV also were significantly correlated (p = 0.0325) with disease-specific survival in patients diagnosed histologically as grade 2. These findings indicate that estimates of MNV are useful for determining the prognosis of transitional cell carcinoma of the upper urinary tract.     

The mutations induced by oxidatively damaged nucleotides, 5-formyl-dUTP and 5-hydroxy-dCTP,in Escherichia coli

The mutational properties of 5-formyl-2'-deoxyuridine 5'-triphosphate (5-CHO-dUTP) and 5-hydroxy-2'-deoxycytidine 5'-triphosphate (5-OH-dCTP), the major oxidatively damaged pyrimidine nucleotides derived from dTTP and dCTP, respectively, were analyzed by an in vivo assay. 5-CHO-dUTP and 5-OH-dCTP were directly incorporated into Escherichia coli , and their mutagenicities were evaluated by the chromosomal lacI forward mutation assay. The mutation frequencies increased, depending on the dose of these damaged nucleotides, indicating that these nucleotides were incorporated into E.coli and acted as mutagens in vivo . The mutagenicities of 5-CHO-dUTP and 5-OH-dCTP were comparable to that of 8-hydroxy-2'-deoxyguanosine 5'-triphosphate, a major form of dGTP oxidative damage. 5-CHO-dUTP induced G.C to A.T, A.T to G.C and G.C to T.A mutations, and 5-OH-dCTP elicited G.C to A.T, A.T to C.G and G.C to T.A mutations.     

Influence of inoculation site of combined oil-adjuvanted vaccine on the antibody response in chickens

In this study, we evaluated the relationship between resistance to insulin-mediated glucose disposal and hematocrit (Hct) and hemoglobin (Hgb) concentrations in 150 normal, healthy volunteers: 100 men and 50 women. Insulin resistance was defined as the steady-state plasma glucose (SSPG) concentration at the end of a 180-minute infusion of somatostatin, insulin, and glucose. Since the steady-state plasma insulin (SSPI) concentrations are similar in all individuals, the SSPG concentrations provide a direct measure of insulin resistance: the higher the SSPG, the more insulin-resistant the subject. The results indicated that SSPG was significantly (P < .001) related to Hct and Hgb in both men and women, with correlation coefficients (r) ranging from 0.38 to 0.43. A series of other variables were also related to Hct and Hgb, including blood pressure, plasma glucose and insulin responses to oral glucose, and plasma triglyceride and high-density lipoprotein (HDL) concentrations. When multiple regression analysis was used to evaluate these relationships, the only variables that were consistently found to be associated with Hct and Hgb were insulin resistance and plasma insulin response to oral glucose. Thus, these results suggest that Hct and Hgb concentrations be added to the cluster of variables related to insulin resistance and compensatory hyperinsulinemia.     

Development of a high activity HDS catalyst for diesel fuel: from basic research to commercial experience

Cosmo Oil and Petroleum Energy Center (PEC) have developed a new hydrodesulfurization (HDS) catalyst of high activity, C-603A, to produce clean diesel fuel, whose sulfur content is less than 0.05 mass%. The preparation of this catalyst combines the use of zeolite technology and impregnation technology to provide excellent HDS activity. C-603A possesses significantly higher activity than conventional Co–Mo/alumina catalyst. Industrial operation with this catalyst has successfully proven its high performance.     

Industrial production of dihomo-γ-linolenic acid by a Δ5 desaturase-defective mutant of Mortierella alpina 1S-4 Fungus

Dihomo-γ-linolenic acid (DGLA)-containing oil (triacylglycerol) was produced by the fungus Mortierella alpina S14, which is a Δ5 desaturase-defective mutant of the arachidonic acid-producing strain 1S-4. Using soy flour as the nitrogen source, S14 produced 8.1 g DGLA/L of culture medium in a 50-L jar fermenter. Shifting the cultivation temperature from 26 to 28°C resulted in reduction of the percentage of DGLA in total fatty acids. Under optimal conditions in a 10-kL industrial fermenter, DGLA production reached 7.0 g/L (percentage of DGLA, 43.9%) at day 12 of cultivation. The other fatty acids were palmitic (18.2%), stearic (7.9%), oleic (7.5%), linoleic (4.4%), γ-linolenic (3.2%), arachidonic (0.4%), and lignoceric (7.7%) acids. This work was presented at the Biocatalysis Symposium in April 2000, held at the 91 st Annual Meeting and Expo of the American Oil Chemists’ Society, San Diego, CA.