Inhibition of N-methyl-N'-nitro-N-nitrosoguanidine-induced carcinogenesis by (-)-epigallocatechin gallate in the rat glandular stomach.

Recently, an epidemiological study showed a lower risk of gastric cancer among people who consume a large amount of green tea. (-)-Epigallocatechin gallate (EGCG), one of the main constituents of green tea, inhibited tumor promotion by teleocidin in a two-stage carcinogenesis experiment with the use of mouse skin. The inhibitory effect of EGCG on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced carcinogenesis of the glandular stomach in rats was examined. The percentage of tumor-bearing rats in the group treated with MNNG plus EGCG was 31%, compared to 62% in the MNNG group. The difference was statistically significant (P < 0.05). To assess the effect of p.o. administration of EGCG, the gastric mucosal cellular kinetics was examined with the use of the bromodeoxyuridine labeling index, ornithine decarboxylase activity, and tissue polyamine levels. The labeling index of the EGCG treatment group decreased significantly (P < 0.05) compared to the EGCG plus MNNG treatment group. The ornithine decarboxylase activity and tissue spermidine levels were also decreased. On the other hand, the tissue putrescine and spermine levels were partly increased. These findings suggest that EGCG inhibits the cellular kinetics of the gastric mucosa during the promotion stage of MNNG-induced gastric carcinogenesis. EGCG may be useful in preventing gastric carcinogenesis. Moreover, EGCG may be applied clinically without any harmful effects and at a low cost.

Streptococcal preparation (OK-432) inhibits development of type I diabetes in NOD mice.

OK-432 (a streptococcal preparation) has been widely used for cancer immunotherapy in Japan. It is the most potent immunomodulator in activating both macrophages and killer T cells and in increasing interleukin 2 production. Two K.E. (Klinische Einheit, clinical unit) of OK-432 were given intraperitoneally to each of 17 female nonobese diabetic (NOD) mice every week from 4-24 wk of age. NOD mice as well as BB rats spontaneously develop type I diabetes. During administration of OK-432, the development of diabetes was inhibited in 17 of 17 mice over the 24-wk observation period, whereas 14 of 17 female NOD mice given physiological saline had developed diabetes by 24 wk of age. At the onset of diabetes, nonfasting blood glucose was 511 +/- 82 mg/dl. Histologic examination showed that in the OK-432-treated NOD mice, 98% of total islets were intact or mildly infiltrated with mononuclear cells, whereas in saline-treated NOD mice, 79% of total islets exhibited severe insulitis. In OK-432-treated NOD mice, both the number of the mononuclear spleen cells and their natural killer cell activity was significantly increased.

Treatment with streptococcal preparation (OK-432) suppresses anti-islet autoimmunity and prevents diabetes in BB rats.

We have recently shown that a streptococcal preparation (OK-432) inhibits insulitis and prevents diabetes in nonobese diabetic (NOD) mice, an animal model of insulin-dependent diabetes mellitus (IDDM). We extended this study to another model of IDDM, namely BB rats. Male and female BB rats were injected weekly with 0.2 mg OK-432 i.p. starting from 5 to 6 wk and continuing through 20 or 30 wk of age. The cumulative incidence of IDDM over 20 wk in the OK-432-treated BB rats (4 of 54, 7.4%) was significantly (P less than .01) lower than that found in the nontreated BB rats (13 of 47, 27.7%). We examined some of these rats as follows. All of the OK-432-treated BB rats tested showed normal glucose levels before and after oral glucose administrations, as did the nontreated and nondiabetic BB rats. Histological examination of pancreatic sections revealed that the OK-432-treated rats retained a greater number of intact islets without infiltration of the mononuclear cells than did the nontreated BB rats. A preliminary in vitro study further demonstrated that the cytotoxic activities of spleen cells against a rat insulinoma cell line, RIN, were suppressed in the OK-432-treated rat. However, the treatment of BB rats with OK-432 showed no suppressive effects in the spleen cell number, the responsiveness of spleen cells to concanavalin A, the populations of OX19+, W3/25+, and OX8+ peripheral blood lymphocytes, or in the titers of cell surface antibody against RIN. These results suggest that a nonimmunosuppressive immunomodulator such as OK-432 may be useful as an agent for immunotherapy of IDDM.

Immunohistochemical staining and activity of ornithine decarboxylase in colorectal cancer.

Using a new anti-human ornithine decarboxylase (anti-hODC) monoclonal antibody, the relationship between the immunoreactivity of ODC and its activity was analyzed in 21 human colorectal cancer tissues, 42 adjacent non-tumorous mucosa specimens, and 10 normal rectal mucosa samples from frozen sections and paraffin-embedded samples. A statistical significant correlation was found between the antibody reaction and the enzymic activity (P < 0.01). The immunohistochemical staining for ODC provides a new and simplified procedure for studying the activity of ODC as compared to previous methods using radioisotopes. It offers the advantages of retrospectively determining the amount of ODC in samples previously embedded in paraffin.

Thymidine phosphorylase expression and effect of doxifluridine: a phase II study.

Doxifluridine (5'-DFUR), an active intermediate metabolite of capecitabine, is converted to 5-fluorouracil by thymidine phosphorylase (TP). We used immunohistochemical staining to investigate the relation between TP expression and 5'-DFUR effects in 40 patients with advanced/recurrent lung metastases from colorectal cancer. Cox regression analysis suggested that TP-positive cancer cells (risk ratio 3.72), were independent factors in survival whereas factors in progression-free survival were TP-positive cancer cells (2.93), and TP-positive stromal cells (0.24). It is suggested that TP expression in cancer cells and in stromal cells are opposite prognostic factors in patients treated with 5'-DFUR.

Dominantly inherited early-onset non-progressive cerebellar ataxia syndrome.

A mother and daughter with suspected dominantly inherited, early-onset, non-progressive cerebellar ataxia syndrome have been reported. A review of the literature and the clinical features of the present cases revealed the nosologic features of this rare disorder, possibly dominant inheritance, floppiness and delayed milestones preceding early-onset mild cerebellar ataxia, non-progressive clinical course, retained or even brisk tendon reflexes without pyramidal tract involvement, normal or slightly delayed intelligence, and occasional nystagmus. Neuroimaging reveals selective involvement of the cerebellum, which is prominent in the vermis and the anterior part of the hemispheres.

Changes in serum LECT 2 levels during the early period of liver regeneration after adult living related donor liver transplantation.

We investigated changes in serum leukocyte cell-derived chemotaxin2 (LECT2) levels between donors and recipients in the early period during liver regeneration following adult living related donor liver transplantation (LRDLT). Five recipients (three women, two men; 37.0 +/- 15.8 years old), all of whom had end-stage liver failure, underwent LRDLT from healthy five donors (two women, three men; 41.6 +/- 14.3 years old) between June 2000 and February 2001. FK506 and methylprednisolone were used as immunosuppressants for recipients. Serum LECT2 levels decreased immediately after both the hepatectomy in all donors and the implantation of liver graft in all recipients. Donors showed a nadir at 3 to 12 hours, increasing at 24 to 48 hours. The nadir in recipients occurred several hours after the donors. The serum LECT2 levels of donors were significantly higher than those of recipients on day 5 (9.5 +/- 5.9 ng/mL vs 3.1 +/- 2.2 ng/mL, P = .04) and on day 7 (9.3 +/- 3.8 ng/mL vs 3.5 +/- 1.1 ng/mL, P = .04). Serum GPT and GOT levels were inversely proportionate to the serum LECT2 levels. The present studies suggest that LECT2 participates in liver regeneration and injury following hepatectomy.

Serum LECT2 level as a prognostic indicator in acute liver failure.

In the present study, we investigated the relationship between serum leukocyte cell-derived chemotaxin2 (LECT2) levels and liver function in patients with acute liver failure, and its use as a prognostic indicator. We studied six acute liver failure patients (two women, four men; 49.8 +/- 20.7 years old) admitted to our hospital in 2002. These patients had diagnoses of fulminant hepatitis due to acute liver failure (1) from congestive heart failure; (2) from portal venous gas, and (3) from postoperative disseminated intravascular coagulation (DIC). We measured serum LECT2, GOT, and GPT levels, the last two being inversely proportionate to the serum LECT2 levels. When the serum GPT levels peaked, the serum LECT2 levels were the lowest. When the liver function recovered, serum LECT2 levels increased. Three of four patients died due to liver failure, one to congestive heart failure. Maximum serum LECT2 levels among the expired group were significantly lower than those among the alive group (0.96 +/- 0.8 ng/mL vs 12.9 +/- 4.3 ng/mL). Serum LECT2 levels may be a prognostic indicator of recovery from liver failure. The present study suggests that in clinical medicine LECT2 participates in regeneration after injury of hepatocytes.

[Electromyographic study on the external urethral sphincter of the male--power spectrum analysis with fast Fourier transformation].

In 23 male patients, the electrical activity of external urethral sphincter muscle was sampled before examination of cystometry (Rest), at first desire to void (FDV), maximum desire to void (MDV) and during urination (Void) by means of electromyography (EMG). Sampling time of each event was 2400 msec. The electrical activity was amplified with a lower limiting frequency of 20 Hz and an upper limiting frequency of 10000 Hz, and recorded on magneto-optical desk. Spectra were obtained using a Hamming window. The action potential of the muscle was quantitatively analyzed and power spectrum of the needle EMGs were analyzed from the magneto-optical disk by a spectrum analyzer using fast Fourier transformation (FFT, Sande-Tukey method). From each power spectrum, mean power frequency (MPF) was obtained from a calculator connected to the spectrum analyzer. Twenty three patients were divided into three groups as follows: 8 patients without any neurological abnormality (normal group), 7 patients with neurogenic bladder showing detrusor-sphincter-dyssynergia (DSD+ group) and 8 patients with neurogenic bladder without detrusor-sphincter-dyssynergia (DSD- group). The results obtained were as follows: 1) The motor unit potentials at Rest had the mean amplitude of 210 +/- 59.4 microV, 329.3 +/- 157.1 microV and 177.6 +/- 132.8 microV in normal group, DSD+ group and DSD- group, respectively. The mean duration were 4.3 +/- 0.2 msec, 5.9 +/- 1.9 msec, and 7.3 +/- 4.5 msec., respectively. The mean phases were 2.8 +/- 0.6, 2.8 +/- 0.5 and 2.5 +/- 0.3, respectively. Statistically there was no difference in amplitude, duration and phase among three groups. 2) In normal group, the power over than -20 dB was distributed from 150 to 220 Hz windows at Rest and FDV. The needle EMGs at MDV showed interference pattern and the power increased over -20 dB in all windows. The power of high frequency area relatively increased. The action potential of the muscle disappeared during urination and the power also disappeared. The average MPF was 103 Hz, 102 Hz, 150 Hz and 98 Hz at Rest, FDV, MDV and Void, respectively. 3) In DSD+ group, the needle EMGs showed interference pattern in MDV and Void, and the power increased only in low frequency area. The average MPF was 104 Hz, 105 Hz, 114 Hz and 120 Hz at Rest, FDV, MDV and Void, respectively. 4) Because of damage to nerves, the action potential of the muscle was difficult to obtain from 3 patients of DSD- group, and the power was not demonstrated.(ABSTRACT TRUNCATED AT 400 WORDS)

[Combination chemotherapy of doxifluridine plus mitomycin C for colorectal lung metastasis--phase II study].

PURPOSE: We conducted a multi-center study to investigate the usefulness of a combination drug therapy with doxifluridine (5'-DFUR) and mitomycin C (MMC) in colorectal cancer patients with lung metastasis. PATIENT AND METHODS: Subjects were advanced/recurrent colorectal cancer patients with lung metastasis, who underwent concomitant drug administration with 533 mg/m2/day of 5'-DFUR orally and 4 mg/m2/day of MMC every 2 weeks intravenously. RESULTS: Of 84 patients registered, 54 patients who were evaluable for tumor response showed results such as: complete response, one; partial response, 4; no change, 30; and progressive disease, 19, corresponding to a response rate of 9.3%. The median survival period of 54 patients was long at 473 days. The median administration days of 5'-DFUR was 201.5 days and the median number of MMC administrations was 14, indicating a long administration period of the combined therapy. The incidence of adverse drug reactions (ADRs) was 37.2% which included thrombocytopenia, 16.7%, and leukocytopenia, 11.5%; only a few ADRs were grade 3 or over. CONCLUSIONS: While combined therapy with 5'-DFUR and MMC resulted in a low response rate, the regimen suggested a survival effect in the patients.

IgG1 is the dominant subclass of antibody against glutamic acid decarboxylase among type 1 diabetes in Japanese.

Autoantibody against glutamic acid decarboxylase (GADA) is a highly sensitive predictor of insulin-dependency in adult diabetic patients as well as young individuals. A considerable number of diabetics who do not reach the insulin-dependent stage have this antibody. Recently, type 1 diabetes has been thought to be caused by T helper 1 (Thl)-type autoimmunity based on studies in non-obese diabetic mice, but it is still difficult to investigate antigen-specific T-cell function in human type 1 diabetes. We therefore assessed an IgG subclass assay for GADA, which should reflect T-helper function against GAD. Sera from 14 type 1 diabetic patients positive for GADA by radioligand binding assay were tested for the IgG subclass of GADA. The assay was based on an enzyme-linked immunosorbent assay, which showed a good correlation with radioligand binding assay. The sera of all but one of the 14 type 1 diabetic patients (93%) were positive for the IgG1 subclass of GADA. The IgG2 and IgG3 subclasses of GADA were also detected in one diabetic patient each who were also positive for IgG1. The IgG4 subclass was not detected in any of the sera we tested. We concluded that IgG1 is the dominant subclass of GADA in Japanese type 1 diabetic patients.

Risk analysis of carcinogenesis in the remnant stomach with measurement of ornithine decarboxylase activity.

Gastric ornithine decarboxylase (ODC) activity was measured as a biomarker of tumor-promoting activity in the remnant stomach of rats and humans. Gastrectomy of Wistar rats utilizing the Billroth I method caused a significantly high induction of ODC, and use of the Billroth II method caused a significantly higher induction of ODC than the Billroth I method. In humans, ODC activity of remnant gastric cancer tissue, normal-appearing mucosa of remnant gastric cancer patient, and remnant gastric mucosa without cancer after the Billroth II method were significantly higher than that of normal gastric mucosa without gastrectomy. ODC activity of remnant gastric mucosa without cancer after the Billroth II method was significantly higher than that after the Billroth I method. Risk of carcinogenesis was high in the remnant stomach, especially after the Billroth II method.

Characteristics of IL-6 and TNF-alpha production by respiratory syncytial virus-infected macrophages in the neonate.

The production of IL-6 and TNF-alpha and the expression of their mRNA were studied with neonatal (cord blood) and adult blood monocyte-derived macrophages (MDM) after in vitro infection with respiratory syncytial virus (RSV). Cord blood MDM exhibited production of high levels of IL-6 within 24 hr after infection. Little or no IL-6 production was detected after 24-48 hr and after in vitro stimulation with inactivated (nonreplicating) virus. Adult blood MDM also produced high levels of IL-6 within 24 hr of RSV infection. Unlike cord blood MDM, adult MDM demonstrated significant activity of IL-6 after 24 hr of infection with live RSV and after exposure to the inactivated virus. The pattern of TNF-alpha production by cord and adult blood MDM after live RSV infection resembled closely the pattern of IL-6 production. Both cell types produced TNF-alpha in the first 24 hr after infection. However, little or no production was observed after 24 hr of infection and after exposure to the inactivated virus. The profile of mRNA expression was similar to the production of IL-6 or TNF-alpha. mRNA expression occurred over a shorter period in cord blood MDM. These observations suggest that inflammatory and immunoregulatory cytokines, such as IL-6 and TNF-alpha, are produced by neonatal as well as previously primed adult macrophages. However, neonatal cells may be less efficient in inducing IL-6 production.

Inhibitory effects and toxicity of green tea polyphenols for gastrointestinal carcinogenesis.

BACKGROUND: Recently, and epidemiologic study showed a lower risk of gastrointestinal carcinogenesis in green tea drinkers. An experiment on two-stage skin carcinogenesis in mice showed that (-)-epigallocatechin gallate (EGCG), one of the main constituents of green tea, inhibited tumor formation. METHODS: The inhibitory effects of EGCG and green tea extract (GTE) on N-ethyl-N'-nitro-N-nitroguanidine (ENNG)-induced duodenal carcinogenesis in the mouse, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced carcinogenesis of the glandular stomach in the rat, and azoxymethane-induced colon carcinogenesis in the rat were examined. The toxicity of GTE was assessed experimentally and GTE was applied clinically in normal volunteers to determine the effective dose and to assess its harmful effects. RESULTS: EGCG and GRE inhibited chemical carcinogenesis of the gastrointestinal tract in rodents. Judging from the epidemiologic and experimental findings, it was determined that 1 g per day of GTE might be an effective dose. GTE was not toxic and no harmful effect was found during its clinical use. CONCLUSIONS: These findings suggest that EGCG and GTE are useful in preventing gastrointestinal carcinogenesis, and the clinical usefulness of GTE, which has no harmful effects and is inexpensive, should be studied further.

Obsessive-compulsive disorder with poor insight.

Although a diagnosis of obsessive-compulsive disorder (OCD) can be made with the specification "poor insight" (PI), this subtype remains understudied. To investigate the subtype, 78 OCD patients were characterized by degree of insight, reevaluated after treatment, and compared with 20 schizophrenics with OCD (OCD+S). At the pretreatment assessments in OCD patients, 28 subjects with poor or delusional insight (PI; 36%) were distinguished from 50 subjects with fair or good insight (GI; 64%) using the insight question of the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS). Compared to the OCD+S group, OCD patients were less likely to have PI, whereas OCD PI patients showed a similar degree of functional impairment as that observed in the OCD+S. After a 6-month combination of clomipramine with cognitive-behavioral treatment, 14 of 25 OCD PI patients no longer fell in the PI category, which was associated with reduced OCD severity and depressive status. Schizotypal personality disorder (SPD) was more common in patients whose insight remained poor even after the treatment. OCD patients demonstrate a range of insight with PI accompanied by significant dysfunction. Comorbid SPD in PI patients may be associated with worse prognosis.

Endogenously generated nitric oxide by nitric-oxide synthase gene transfer inhibits cellular proliferation.

We investigated whether endothelial nitrite oxide synthase (NOS) gene transfer inhibited cellular proliferation. Endothelial NOS and endothelin type A receptor genes were transferred into 293 cells, a human embryonic kidney cell line, by calcium-phosphate coprecipitation. The cytosolic free Ca(2+) levels ([Ca(2+)](i)) of transfected cells were estimated with fura-2 fluorescence. Thymidine incorporation was increased by endothelin-1 in type A receptor-transfected cells. The endothelial NOS gene transfer did not affect endothelin-1-induced increase in [Ca(2+)](i) of type A receptor-transfected cells, but markedly inhibited mitogen-activated protein kinase and c-fos promoter activities. The endothelial NOS gene transfer also inhibited thymidine incorporation into type A receptor-transfected cells in response to endothelin-1, which was abolished in the presence of the NOS inhibitor N(G)-monomethyl-L-arginine acetate. The endothelin-1-induced increase in cell number was significantly suppressed by endothelial NOS gene transfer as well as by the mitogen-activated protein kinase inhibitor PD98059. These results indicate that endothelial NOS gene transfer inhibits cellular proliferation via inhibition of the mitogen-activated protein kinase cascade.