Multicenter randomized phase II study of cisplatin and fluorouracil plus docetaxel (DCF) compared with cisplatin and fluorouracil plus Adriamycin (ACF) as preoperative chemotherapy for resectable esophageal squamous cell carcinoma (OGSG1003).

BACKGROUND: This phase II trial evaluated the efficacy of cisplatin and fluorouracil (CF)-based combination neoadjuvant chemotherapy on the outcome of patients with resectable locally advanced esophageal squamous cell carcinoma (ESCC). We compared the recurrence-free survival (RFS) associated with CF plus Adriamycin (ACF) with that associated with CF plus docetaxel (DCF) to select an alternative regimen in a new phase III trial investigating the optimal neoadjuvant treatment of patients with ESCC. PATIENTS AND METHODS: Patients with resectable advanced ESCC were randomly assigned to either ACF (Adriamycin 35 mg/m2, cisplatin 70 mg/m2 i.v. on day 1, fluorouracil 700 mg/m2 continuous infusion for 7 days) every 4 weeks or DCF (docetaxel 70 mg/m2, cisplatin 70 mg/m2 i.v. on day 1, fluorouracil 700 mg/m2 continuous infusion for 5 days) every 3 weeks. Surgery was scheduled after completion of two cycles of chemotherapy. The primary end point was RFS, analyzed by the intention-to-treat. RESULTS: Between October 2011 and October 2013, 162 patients at 10 institutions were enrolled in the study, all of whom were eligible and randomly assigned to the two groups (81 to the ACF group and 81 to the DCF group). The R0 resection rates for the ACF and DCF groups were equivalent (95.9% versus 96.2%, P = 0.93). The 2-year RFS and overall survival rates for DCF versus ACF were 64.1% versus 42.9% (hazard ratio 0.53, 95% confidence interval 0.33-0.83, P = 0.0057) and 78.6% versus 65.4% (P = 0.08), respectively. CONCLUSION: Compared with ACF, DCF chemotherapy was associated with prolonged RFS for patients with resectable advanced ESCC. Thus, DCF chemotherapy has potential as a standard neoadjuvant therapy for resectable ESCC. CLINICAL TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trials Registry of Japan (identification number UMIN000004555/000004616).

Experience of a specialist centre in the management of anastomotic sinus following leaks after low rectal or ileal pouch-anal anastomosis with diverting stoma.

AIM: The natural history and appropriate management of anastomotic sinus has not been clearly defined. The aim of this study was to evaluate the incidence, management and outcomes of anastomotic sinus. METHOD: The medical records of all patients who underwent a low anterior resection (LAR) or an ileal pouch-anal anastomosis (IPAA) with a diverting loop ileostomy (LI) and with contrast enema performed before planned stoma closure between 2001 and 2011 were retrospectively reviewed. The radiological features of the sinus tract, treatment and outcome of anastomotic sinus were studied. RESULTS: Twenty patients (8.2%) were found to have anastomotic sinuses out of the total of 244 patients who had undergone LAR (n = 146) or IPAA (n = 98) with LI. Of these, 13 (65%) had prior symptomatic leaks, while seven did not. Twelve patients (60%) were found to have simple sinus tracts, while eight had complex sinuses (associated with either pelvic cavities or severe strictures). Five patients with simple tracts were treated with observation alone. Fifteen patients underwent surgical interventions. Overall, with a median follow-up of 28 (6-73) months, 16 patients (80%) had resolution of their sinuses. All of 12 patients (100%) with simple sinus tracts and four of eight patients (50%) with complex sinuses underwent successful stoma reversals after 8 (3.5-24) months following the initial surgery (P = 0.01). CONCLUSION: Patients with simple tracts are significantly more likely to have complete resolution of sinuses than patients with complex sinuses. Persistent sinus associated with either a pelvic cavity or severe stricture despite surgical intervention is likely to lead to a permanent stoma.

Role of multidrug resistance protein 2 (MRP2) in chemoresistance and clinical outcome in oesophageal squamous cell carcinoma.

BACKGROUND: Although multidrug resistance protein 2 (MRP2) confers chemoresistance in some cancer types, its implication on oesophageal squamous cell carcinoma (ESCC) remains unclear. METHODS: We evaluated MRP2 expression by immunohistochemistry and RT-PCR using 81 resected specimens from ESCC patients who did or did not receive neo-adjuvant chemotherapy (NACT), including 5-fluorouracil, doxorubicin, and cisplatin (CDDP). Correlation between MRP2 expression and response to chemotherapy was also examined in 42 pre-therapeutic biopsy samples and eight ESCC cell lines. RESULTS: MRP2-positive immunostaining was more frequently observed in ESCCs with NACT than in those without NACT (27.3 vs 5.4%). The MRP2-positive patients showed poorer prognosis than MRP2-negative patients (5-year survival rate, 25.6 vs 55.7%). Concordantly, ESCC with NACT showed 2.1-fold higher mRNA expression of MRP2 than those without NACT (P=0.0350). In pre-therapeutic biopsy samples of patients with NACT, non-responders showed 2.9-fold higher mRNA expression of MRP2 than responders (P=0.0035). Among the panel of ESCC cell lines, TE14 showed the highest MRP2 mRNA expression along with the strongest resistance to CDDP. Inhibition of MRP2 expression by small-interfering RNA reduced chemoresistance to CDDP. CONCLUSION: Our data suggested that MRP2 is one of molecules, which regulate the sensitivity to chemotherapy including CDDP in advanced ESCC patients.

Dietary intake estimations of polybrominated diphenyl ethers (PBDEs) based on a total diet study in Osaka, Japan.

This study presents the results of a total diet study performed for estimating the dietary intake of polybrominated diphenyl ethers (PBDEs) in Osaka, Japan. The concentrations of 36 PBDEs were measured in samples from 14 food groups (Groups I-XIV). PBDEs were detected only in Groups IV (oils and fats), V (legumes and their products), X (fish, shellfish, and their products), and XI (meat and eggs) at concentrations of 1.8, 0.03, 0.48, and 0.01 ng g⁻¹, respectively. For an average person, the lower bound dietary intakes of penta- and deca-formulations were estimated to be 46 and 21 ng day⁻¹, respectively. A high proportion of the decabrominated congener (DeBDE-209) was observed in Group IV. To confirm the presence of DeBDE-209 in vegetable oils, an additional analysis was performed using 18 vegetable oil samples. Of these, seven contained ng g⁻¹ levels of DeBDE-209.

Effect of oral treatment of Perilla frutescens and its constituents on type-I allergy in mice.

Perilla frutescens Britton (perilla, Labiatae) is a medicinal herb prescribed in Saiboku-to [Japanese letters: see text], which is a Kampo formula effective for allergic diseases such as bronchial asthma. The present study was conducted to evaluate the anti-allergic effect of orally administered perilla decoction and to identify the active constituents using mice ear-passive cutaneous anaphylaxis (PCA)-reaction, which is one of the animal models for type I allergy. Perilla decoction significantly suppressed PCA-reaction, and the inhibition % at the dose of 500 mg/kg was 43%. The perilla decoction contains 5.3% of luteolin 7-O-[beta-glucuronosyl(2-->1)beta-glucuronide], 1.6% of apigenin 7-O-[beta-glucuronosyl(2-->1)beta-glucuronide], 0.49% of scutellarin, and 2.5% of rosmarinic acid (weight of compound/dried weight of perilla decoction %), respectively. When these constituents were orally administered to mice at the dose equivalent to 500 mg/kg of perilla decoction, rosmarinic acid and apigenin 7-O-[beta-glucuronosyl(2-->1)beta-glucuronide] significantly suppressed PCA-reaction, and their inhibition % was 41% (p<0.01) and 32% (p<0.05), respectively. Since the inhibition % or perilla decoction and rosmarinic acid were nearly equal, the anti-allergic effect of perilla decoction depends primarily on rosmarinic acid. The standard Saiboku-to decoction contained 0.013% of rosmarinic acid, which was too low to exhibit anti-allergic activity in a daily dose of Saiboku-to in adults, suggesting that perilla would be prescribed in Saiboku-to to exhibit other pharmacological effects than its anti-allergic activity, such as a sedative.

Hornerin, a novel profilaggrin-like protein and differentiation-specific marker isolated from mouse skin.

A novel mouse cDNA named hornerin was isolated by RNA differential display applied to developing mouse skin. Hornerin, which has 2,496 amino acids, comprises EF-hand domains at the N terminus followed by a spacer sequence and a large repetitive domain, indicating that hornerin is a novel member of the "fused gene"-type cornified envelope precursor protein family. The repetitive domain of hornerin was found to be rich in glycine, serine, and glutamine. Hornerin was expressed in the tongue, esophagus, forestomach, and skin among the adult mouse tissues examined, all of them cornifying stratified epithelium. In the embryonic mouse skin, hornerin mRNA was first detected on gestational day 15.5 in the epidermis coincidentally with the formation of a granular layer. In accordance with this, hornerin was detected in the granular and cornified layers of the mature epidermis. In the granular cells of the epidermis, the hornerin protein was detected in keratohyalin granules together with profilaggrin. Furthermore, Western blot analysis of the mouse skin showed that the hornerin protein was cleaved during the process of epidermal differentiation, indicating possible posttranslational proteolytic processing as is observed in profilaggrin. Differentiation of primary mouse epidermal keratinocytes with 0.12 mm Ca(2+) resulted in the induction of hornerin. These results indicate that hornerin is structurally as well as functionally most similar to profilaggrin among the family members and possibly plays pleiotropic roles, including a role in cornification.

Presence of aldose reductase inhibitors in tea leaves.

Water extract from commercial English tea has a potent inhibitory activity against human placenta aldose reductase (NADPH oxidoreductase, E.C.1.1.1.21.). Inhibitory activity was separated into five major fractions by one-step chromatography with a C-18 reverse phase column. The most active fraction was further subjected to reverse phase column chromatography. As a result, a well-known flavone-glycoside, isoquercitrin, was isolated as the most potent chemical. The inhibitory character of isoquercitrin for aldose reductase was a mix of uncompetitive and noncompetitive inhibitions, and its IC50 was 1 x 10(-6) M. In rat sciatic nerve tissue preparations, sorbitol accumulation in the presence of high concentrations of glucose (30 mM) was inhibited by 38% at 5 x 10(-4) M of isoquercitrin. The flavone-glycoside isoquercitrin is the active inhibitor of aldose reductase inhibitor present in English tea. Given the ability of aldose reductase inhibitors to prevent diabetic complications, an epidemiological study of the effect of tea consumption on the pathogenesis and progression of diabetic complications would be interesting.

Inhibitory effect of decoction of Perilla frutescens on cultured murine mesangial cell proliferation and quantitative analysis of its active constituents.

The leaves of Perilla frutescens (perilla) are a common herb used in Japan for garnishing raw seafood. Previously, we reported that a decoction of perilla leaves had suppressive effects on the progression of glomerulonephritis in an animal model of spontaneous IgA nephropathy. The objective of the present study was to isolate anti-nephritic constituents in the perilla decoction under the guidance of its in vitro anti-proliferative activity on cultured murine mesangial cells, and to measure the contents of the active constituents in decoctions prepared from various perilla chemotypes, which differ in their composition of essential oils and/or pigments. DNA synthesis of cultured mesangial cells induced by 1% fetal calf serum was significantly inhibited by the perilla decoction (IC50 values, 8.8 microg/ml). Caffeic acid, luteolin 7-O-[beta-glucuronosyl(1-->2)beta-glucuronide], apigenin 7-O-[beta-glucuronosyl(1-->2)beta-glucuronide], scutellarin, and rosmarinic acid were isolated as active constituents. The contents of these phenolic compounds were not significantly different among chemotypes of P. frutescens. Considering the relation between the contents in the perilla decoction and the activities of these compounds, rosmarinic acid represents the in vitro anti-proliferative effect of perilla decoction.

Suppressive effects of Perilla frutescens on mesangioproliferative glomerulonephritis in rats.

Leaves of Perilla frutescens var. crispa DECNE. (perilla, Labiatae) are used as a garnishing vegetable in East Asian countries as well as an herbal medicine prescribed in Kampo medicines such as Saiboku-to. A previous in vitro study revealed that a decoction of perilla leaves inhibits the proliferation of murine-cultured mesangial cells. In the present study, we evaluated the in vivo anti-proliferative effects of a perilla decoction using rat mesangio-proliferative glomerulonephritis induced by an intravenous injection of rabbit anti-rat thymocyte serum (ATS). Leaves of perilla were boiled, and the decoction was orally administered to the rats as drinking water at doses of 100 and 500 mg/kg/d from the day of ATS-injection (day 0) to day 8, when rats were sacrificed. In the histological evaluation, the total number of glomerular cells, proliferating cell nuclear antigen (PCNA) positive cells, and macrophage/monocyte antigen-positive cells in the glomerulus, was significantly decreased in perilla-treated rats. A significantly lower level of proliferation was induced by the serum of the perilla-treated rats than by that of the controls. These results suggest that the perilla decoction suppresses the proliferation of mesangial cells in vivo by an inhibition of the glomerular infiltration of macrophage/monocytes and of the production of circulating growth factors.

Production of menaquinones by lactic acid bacteria.

Lactic acid bacteria were examined for their ability to produce quinone compounds, which may include dietary sources of menaquinones. Isoprenyl quinones in bacterial cells grown in a synthetic medium were extracted and analyzed by thin layer chromatography. Lactococcus lactis ssp. cremoris (three strains), Lactococcus lactis ssp. lactis (two strains), and Leuconostoc lactis were selected as high producers of quinone that synthesized more than 230 nmol of quinones/g of dried cells. The quinones were presumed to be menaquinone-7 to -10 by high performance liquid chromatography. Precise molecular weights were determined by mass spectrometry for Lactococcus lactis ssp. cremoris YIT 2011 and Leuconostoc lactis YIT 3001 and identified as menaquinone-8 and -9 for the former and menaquinone-9 and -10 for the latter. Those strains, when grown either in reconstituted nonfat dry milk or a soymilk medium, produced a beneficial quantity for dietary supplement (i.e., 29 to 123 micrograms of menaquinones/L of the fermented medium).

Suppressive effects of Perilla frutescens on spontaneous IgA nephropathy in ddY mice.

Perilla frutescens (perilla) is a common herb used in Japan for garnishing raw seafood to protect the alimentary tract from inflammatory diseases. The present study was performed to investigate whether or not perilla prevents the development of lesions of IgA nephropathy in ddY mice which spontaneously develop this disease. After orally administering perilla extract to ddY mice from 8 to 42 weeks of age, the changes in urine, serum, and kidneys were evaluated. Perilla extract significantly suppressed proteinuria and glomerular IgA deposition (p < 0.01 and p < 0.05, respectively). The decreased serum IgA concentration in perilla-treated mice showed a significant correlation with glomerular IgA deposition. Such findings suggest that perilla reduced glomerular IgA deposition via suppression of IgA production in the serum. On the other hand, the nitric oxide concentration in the serum of perilla-treated mice was significantly higher than that observed in the controls. The addition of the sera of perilla-treated mice to quiescent cultured murine mesangial cells resulted in a cell proliferation which was less than in controls, suggesting that perilla might either directly prevent mesangial cell proliferation or prevent proliferation by regulating circulating cytokines. Such results indicate that perilla should prevent IgA nephropathy, thus representing a promising herbal medicine for glomerulonephritis.

Effect of Perilla frutescens extract on nitric oxide production by cultured murine mesangial cells.

The effects of a water extract of perilla (Perilla frutescens Britton) leaves on nitric oxide (NO) production by cultured murine mesangial cells were investigated. Perilla extract significantly induced NO production from mesangial cells, which was enormously augmented without cytotoxity by combination with interferon (IFN)-gamma or tumor necrosis factor-alpha. On the other hand, perilla extract suppressed a large amount of NO production induced by IFN-gamma combined with lipopolysaccharide. Northern blot analysis revealed that such effects of perilla extract were dependent on inducible NO synthase mRNA expression. Perilla extract exhibited an inhibitory effect on cytokine-induced mesangial cell proliferation, and this effect was significantly decreased upon combination with N(G)-monomethyl-L-arginine (L-NMMA), a non-specific NO synthase inhibitor, suggesting that perilla extract inhibits mesangial cell proliferation partially through the induction of NO production. Such results indicate that perilla may be a promising agent for the prevention of the progression of glomerulonephritis.

Inhibitory effect of Perilla frutescens and its phenolic constituents on cultured murine mesangial cell proliferation.

The inhibitory effects of Perilla frutescens and its phenolic constituents on cytokine-induced proliferation of murine cultured mesangial cells were investigated. DNA synthesis of mesangial cells stimulated by platelet derived growth factor (PDGF, 10 ng/ml) or tumor necrosis factor (TNF)-alpha (100 U/ml) was inhibited by the extract of P. frutescens (IC50 values, 3.3 and 1.4 micrograms/ml, respectively). The strength of the anti-proliferative activity was nearly equal in various chemotypes of P. frutescens. Caffeic acid, methyl caffeate, rosmarinic acid, and luteolin 7-O-glucuronide-6"-methyl ester were isolated as active constituents from the extract of the typical strain of P. frutescens, and their IC50 values for PDGF-induced mesangial cell proliferation were estimated as 26 microM, 2.6 microM, 1.8 microM, and 4.1 microM, respectively. We also compared the activities of related flavonoids previously isolated from P. frutescens, and luteolin had the highest anti-proliferative activity.

Pre-operative radio-chemotherapy enhances apoptotic cell death in oral squamous cell carcinoma.

The effect of pre-operative radio-chemotherapy (RCT) has been examined in a total of 15 oral squamous cell carcinomas (SCCs), in terms of apoptosis (cell loss) and proliferation. All the patients received pre-operative radiation at a dosage of 30 or 40 Gy, as well as anticancer agents including tagaful (FT), 5-fluorouracil (5-FU), bleomycin (BLM) and peplomycin (PEP). Surgical specimens were obtained before and after RCT, and serial sections were prepared for immunohistochemistry for p53 oncoprotein and Ki-67 antigen, as well as for terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL). TUNEL indices (TI; percentage of TUNEL-positive cells in the tumor cells) before and after RCT were 1.2+/-1.1 and 4.7+/-2.9 in the nine well-differentiated oral SCCs, and 1.0+/-0.7 and 3.9+/-2.1 in the six poorly differentiated SCCs, respectively. Similarly, Ki-67 indices (KI; percentage of Ki-67 antigen-positive cells in tumor cells) before and after RCT were 31.1+/-14.2 and 15.8+/-11.1 in the former, and 37.1+/-7.8 and 8.7+/- 13.4 in the latter, respectively. Thus, pre-operative RCT enhanced apoptotic cell death and abated proliferative activity significantly (P<0.05), regardless of histological differentiation. Enhancement of apoptosis was more prominent in the group treated with FT or 5-FU than with BLM or PEP. Oral SCC with >20% of nuclear p53-positive tumor cells was noted in six cases. Enhanced TI and abadement of KI did not differ among the p53-positive and -negative tumors.

Anti-allergic constituents in the culture medium of Ganoderma lucidum. (I). Inhibitory effect of oleic acid on histamine release.

The chloroform extract from Ganoderma lucidum broth markedly inhibited histamine release from rat peritoneal mast cells. From the active fractions, palmitic acid, stearic acid, oleic acid and linoleic acid were isolated. Oleic acid dose-dependently inhibited the histamine release and 45Ca uptake into mast cells induced by compound 48/80 and A-23187 at concentrations of 5 to 50 microM and 0.5 to 5 microM, respectively. Saturated fatty acids, however, had only a weak inhibitory effect on histamine release. Although linoleic acid and linolenic acid effectively prevented this release, these two compounds caused marked release at concentrations higher than 10 microM and 20 microM, respectively. Oleic acid induces membrane-stabilization in model membrane systems. It was concluded that one of the effective constituents obtainable from the chloroform extract of G. lucidum-cultured broth is oleic acid.

Anti-allergic constituents in the culture medium of Ganoderma lucidum. (II). The inhibitory effect of cyclooctasulfur on histamine release.

For centuries, Ganoderma lucidum has been used in Oriental medicine for the treatment of chronic bronchitis. Sequential fractions of the culture medium of this plant revealed that one of the active constituents was cyclooctasulfur. The latter effectively inhibited histamine release from rat peritoneal mast cells and impeded 45Ca uptake into these cells without affecting the cyclic AMP content. SDS-PAGE analysis indicated that cyclooctasulfur induced some changes in protein bands obtained from the membrane fraction of mast cells, suggesting that this compound interacts with membrane proteins so as to inhibit 45Ca uptake, and that this may be the main cause of histamine release inhibition.

Self-priming effect of LH-RH on pituitary gonadotropins in hyperprolactinemic women.

To investigate how various concentrations of serum prolactin (PRL) influence the priming effect of luteinizing hormone releasing hormone (LH-RH) on the pituitary gland, 24 women with various blood PRL concentrations received intravenous injections of 100 micrograms of synthetic LH-RH twice at an interval of 60 minutes and their serum LH and follicle-stimulating hormone (FSH) were measured and analysed. In the follicular phase with a normal PRL concentration (PRL less than 20 ng/ml, n = 6), marked first peaks of the two hormones following the first LH-RH stimulation and enhanced second peaks after the second LH-RH administration were observed, indicating a typical priming effect of LH-RH on gonadotropins, though the second response of FSH was more moderate than that of LH. In hyperprolactinemia, in which the serum PRL concentration was higher than 70 ng/ml (n = 13), the basal concentration of gonadotropins was not significantly changed but the priming effect of LH-RH on LH and FSH was significantly decreased (p less than 0.01). No marked second peaks of LH and FSH were observed, suggesting an inhibitory effect of hyperprolactinemia on the second release of LH and FSH. In contrast, this effect was restored in a group of women whose serum PRL concentration was between 30 and 50 ng/ml (n = 5). Furthermore, enhanced second peaks of both LH and FSH were noted after successful bromocriptine therapy reduced hyperprolactinemia (PRL greater than 70 ng/ml) to less than 25 ng/ml (n = 5).(ABSTRACT TRUNCATED AT 250 WORDS)

[Study on the development of the radioimmunoassay of the corticotropin releasing factor (CRF)].

Synthetic ovine CRF was conjugated to bovine serum albumin (BSA) with glutaraldehyde and used as an immunogen to generate antibodies in rabbits. One of the antisera revealed 40% binding with 125I-CRF at 1 : 12,000 dilution in RIA. The antiserum did not show any significant crossreaction with other hormones. The Chloramine-T method was used to label CRF with 125I and purified by Sephadex G-50 fine (1 X 103 cm) chromatography with 0.03 M phosphate buffer (pH 7.2) containing 0.1% BSA, 20mM EDTA, 200 KIU/ml Trasyrol was used for the RIA buffer and the separation of free and antibody-bound CRF was performed by the Dextran-Charcoal method. The tentative minimum sensitivity of the assay was 20-100 pg/tube. The dilution curve of 125I-CRF in the hypothalamus of rat was parallel to that of the standard. Immunoreactive CRF content in the hypothalamus of male, female and post ovariectomized rats was measured by RIA. The content in those of male and post ovariectomized rats was significantly higher than in female rats. These data indicate that the pituitary-adrenal axis may have some effects on post-menopausal status.

Time course of effects of morphine on hypothalamic content of LHRH and serum testosterone and LH levels of morphine-tolerant and nontolerant male rats.

Time course of the effects of morphine on the hypothalamic content of LHRH in both nontolerant and morphine-tolerant male rats was investigated in relation to the temporal changes of serum testosterone and LH levels. The hypothalamic LHRH content of the nontolerant rats was increased 8 hr after the administration of morphine (100 mg/kg) when serum testosterone levels were depressed. The LHRH content of the tolerant rats was decreased during withdrawal of morphine for 48 hr when the lowered serum testosterone and LH levels had returned to within the control levels. Although the hypothalamic LHRH content does not necessarily reflect the release of LHRH, these results are in favour of the hypothesis that the release of hypothalamic LHRH is inhibited by the administration of morphine and is restored by the withdrawal of the narcotic.