15-Deoxy-Delta12,14-prostaglandin J2 inhibits IL-1beta-induced cyclooxygenase-2 expression in mesangial cells

BACKGROUND: Cyclooxygenase-2 (COX-2), a key enzyme in the synthesis of prostaglandins, is induced in mesangial cells in response to proinflammatory cytokines. Recently, 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2), one of the natural ligands of peroxisome proliferator-activated receptor gamma (PPARgamma), has been reported to have an anti-inflammatory effect. Therefore, we examined the effect of 15d-PGJ2 on COX-2 expression in cultured rat mesangial cells.METHODS: Mesangial cells were incubated with 15d-PGJ2 for 30 minutes and then exposed to interleukin-1beta (IL-1beta). The expression of COX-2 mRNA and proteins was determined by Northern blot and immunoblot analyses, respectively. Accumulation of prostaglandin E2 (PGE2) was measured by an enzyme-linked immunosorbent assay (ELISA). Activities of mitogen-activated protein kinases (MAPKs) were evaluated by an immunoblot analysis. DNA binding activities of activator protein-1 (AP-1) or nuclear factor-kappaB (NF-kappaB) were examined by an electrophoretic mobility shift assay (EMSA). The activities of PPAR responsive elements (PPRE) and COX-2 promoter were measured by a luciferase reporter assay.RESULTS: 15D-PGJ2 significantly suppressed IL-1beta-induced COX-2 expression and PGE2 production, but thiazolidinediones, synthetic PPARgamma ligands, did not affect COX-2 expression. Moreover, the cells transfected with a PPRE luciferase reporter did not respond to 15d-PGJ2. IL-1beta rapidly activated extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK), which were involved in the up-regulation of COX-2 induction, but 15d-PGJ2 inhibited the activation of these kinases. 15d-PGJ2 inhibited the IL-1beta-induced increase in binding activities of nuclear proteins to consensus AP-1 site and AP-1-like site of COX-2 promoter but not of NF-kappaB. IL-1beta was unable to activate the COX-2 promoter when the AP-1-like site was mutated.CONCLUSIONS: These data suggest that 15d-PGJ2 inhibits IL-1beta-induced COX-2 expression, independent of PPARgamma activation, by suppression of ERK and JNK pathways and AP-1 activation in mesangial cells. Thus, 15d-PGJ2 may play an important role in the negative feedback mechanism of COX-2 expression in renal inflammation and may be useful as an anti-inflammatory agent.     

A new regimen for TS-1 therapy designed to minimize adverse reactions by introducing a one-week interval after each two-week dosing session

It has been reported that the response rate to TS-1 of advanced recurrent gastric cancer was the highest rate (46.5%) of effectiveness among anti-cancer agents, but the incidence of adverse reactions to this drug has been found to be as high as 83.2%, with grade 3 or severer reactions occurring in 20.3% of patients. Taking into consideration the post-marketing survey finding that adverse reactions to the drug first appear 2-3 weeks after the start of oral TS-1 therapy, we attempted a new dosing regimen for this drug, wherein each session of therapy lasted for 2 weeks, with a one-week interval between two consecutive sessions (herein-after called "the 2-week regimen"). This regimen was employed based on the expectation that the adverse reactions to the drug would be minimized and that the consecutive dosing period could be prolonged, while keeping the anti-cancer potency at a level similar to that expected with the 4-week dosing regimen with a 2-week interval between sessions (the 4-week regimen). The subjects were 38 patients with advanced or recurrent stomach cancer who were treated with TS-1 at our center between September 1999 and November 2001. Twenty-four patients treated using the 4-week method until January 2001 were taken as a historical control, and compared with 14 patients treated using the 2-week method from February 2001 and afterwards. The incidence of adverse reactions was 71% in the 2-week regimen group against 92% in the 4-week regimen group. The incidence of grade 3 or severe adverse reactions was 8% in the 2-week group and 21% in the 4-week group. Thus, the incidence of adverse reactions was lower in the 2-week group. The percentage of patients who complied with the dosing instructions completely during a 6-month period, as evaluated by the Kaplan-Meier method, was 86% in the 2-week group and 58% in the 4-week group. The response rate, as calculated in patients whose lesions could be evaluated, was 25% in the 2-week group and 19% in the 4-week group. These results suggest that the 2-week regimen may allow safer outpatient drug therapy using TS-1 and merits a trial when considering the QOL of patients. We propose conducting a phase-II multi-center clinical study of this regimen in the near future.     

Meige syndrome with apraxia of lid opening after the discontinuation of sulpiride treatment

We report a case of Meige syndrome with apraxia of lid opening that lasted for about seven months after discontinuation of sulpiride treatment. To our knowledge, this is the first report demonstrating that Meige syndrome with apraxia of lid opening is induced by sulpiride, and that the condition persists.     

Utility of follow-up studies using meta-[123 I]iodobenzylguanidine scintigraphy for detecting recurrent neuroblastoma

Neuroblastomas sometimes recur after the initial disappearance of the tumour. We evaluated the utility of meta-[123I]iodobenzylguanidine (123I-MIBG) scintigraphy for the detection of recurrent neuroblastomas by comparing with the measurement of biochemical markers and clinical findings. Forty patients who had received treatment for neuroblastomas were included in the study. After the disappearance of the initial tumours, periodic measurements of urinary vanillyl mandelic acid, homovanillic acid and serum neuron specific enolase values, and an 123I-MIBG scintigraphy were performed. Whenever an abnormal finding was observed, other appropriate examinations and/or follow-up examinations were performed to elucidate the true state of the patient. Eleven recurrent episodes in eight patients were observed. Most of them occurred in the bone marrow or bone. Corresponding symptoms were observed in only two episodes; the other episodes were asymptomatic, and discovered by the periodic examinations. 123I-MIBG scintigrams visualized the recurrent tumours in 10 (91%) episodes. Elevated tumour markers were observed in only three episodes. 123I-MIBG scintigrams visualized most of the recurrent tumours, unless they were accompanied by any symptoms or elevations in biochemical tumour markers. Periodic examinations with 123I-MIBG scintigraphy appears to be a useful technique for the detection of the recurrences.     

Immunocytochemical localization of a neuron-specific thrombospondin-1-like protein, NELL2: light and electron microscopic studies in the rat brain

We have studied the cellular and intracellular localization of NELL2, a neural thrombospondin-1-like protein. NELL2 protein was detected as doublet bands of 140 and 90 kDa with the use of the specific antibodies raised against the C-terminal region of NELL2 and was recognized only in the brain but not in the peripheral tissues. Within the brain, NELL2 was abundantly present in the hippocampus and cerebral cortex, found moderately in the olfactory bulb and hypothalamus, and at a low level in the thalamus, cerebellum, and medulla. Immunocytochemically, NELL2 was seen only in neurons but not in glial cells or in the white matter. NELL2-immunoreactive cells were distributed throughout the brain with the highest density in the hippocampus and cerebral cortex. NELL2 was mainly found in the cell bodies of neurons and the immunoreactivity was often seen as dots in the perikarya. The distribution of NELL2 immunoreactivity did not completely correspond to that of any subtypes of protein kinase C (PKC). Under electron microscopy, NELL2 protein was associated with the endoplasmic reticulum (ER), especially with rough ER. NELL2 immunoreactivity was found in the restricted parts of the ER and found commonly inside the ER. These results suggest that NELL2 protein is synthesized by neurons and may be secreted from the neurons involved in certain neuronal functions.     

Suppression of Lung Metastasis by Aspirin but Not Indomethacin in an in vivo Model of Chemically Induced Hepatocellular Carcinoma

To examine the effect of non-steroidal anti-inflammatory drugs on metastasis formation, aspirin (ASP, 0.5% in diet) and indomethacin (IM, 0.005% in drinking water) were applied to an in vivo highly metastatic rat hepatocellular carcinoma (HCC) model in F344 male rats. Administration for 8 weeks after induction of highly metastatic HCC by sequential treatment with diethylnitrosamine and N-nitrosomorpholine did not cause any significant change in survival rate or body weight. Multiplicity of HCC in the liver increased during ASP or IM treatment without any significant histological alteration. Although absent in the rats killed at the end of the period of carcinogen exposure, lung metastasis at the end of the experiment was found in 100%, 89% and 100% of rats in the control, ASP and IM groups, respectively. Degree of metastasis was classified into three groups according to the number of metastatic nodules, i.e., slight (1–5 nodules), moderate (6–50) and severe (more than 51), which amounted to 0%, 43% and 57% in the control group. ASP significantly reduced the degree of metastasis, the incidences being 33%, 44%, and 11%, respectively, whereas IM was without significant influence. Both agents suppressed cell proliferation in HCCs, without any alteration of pan-cadherin expression. However, expression in HCC of mRNAs for intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, both of which are considered to play key roles in attachment of cancer cells to the endothelium, was significantly suppressed by ASP. Thus, the present study demonstrated that ASP, but not IM, has the potential to inhibit lung metastasis of rat HCC in vivo , possibly via reduced attachment of tumor cells to the vascular endothelium. Moreover, these data indicate this in vivo model for induction of rat highly metastatic HCC to be a useful tool for the assessment of the efficacy of therapeutic treatments to block metastasis formation.     

Prediction of non-responsiveness to intravenous high-dose gamma-globulin therapy in patients with Kawasaki disease at onset

Children with Kawasaki disease (n = 82), treated with intravenous immune globulin (IVIG) at a high dose, were classified as IVIG-responsive (defervescence within 5 days of starting IVIG, n = 69) or IVIG-non-responsive (consistent fever over a 6-day period since starting IVIG, n = 13). One patient in the IVIG-responsive group had a coronary artery abnormality during the acute phase (1. 4%) versus 5 in the IVIG-non-responsive group (38.5%). Age, duration of fever before the initiation of IVIG therapy, and laboratory data obtained on admission were tested by the Mann-Whitney U test. Serum levels of C-reactive protein, total bilirubin, lactate dehydrogenase, and gamma-glutamyltranspeptidase were significantly higher (P =.002, P <.001, P <.034, and P <.038, respectively), and the hemoglobin value was significantly lower (P =.025) in patients in the non-responsive group. A multivariate analysis showed that serum levels of C-reactive protein (P =.006), lactate dehydrogenase (P =. 035), and total bilirubin (P =.046) on admission were independent correlates of the success of IVIG therapy. By defining the predictive values, patients with a C-reactive protein level >10 mg/dL, LDH level >590 IU/L, and/or hemoglobin value <10 g/dL are considered non-responsive to IVIG. Additional therapy at an early stage of the disease should be considered for patients who are predicted to be IVIG-non-responsive.     

Detection of human papillomaviruses from histologically normal lymph nodes of Japanese cervical cancer patients by nested polymerase chain-reaction assay

To determine the prognostic significance of human papilloma virus (HPV) DNA in histologically normal lymph nodes, we developed a nested polymerase chain-reaction (PCR) method on HPV16, 18 and 33 DNAs for formalin-fixed, paraffin-embedded tissues. We investigated 370 histologically normal lymph nodes from 15 patients treated for stage-IB/IIB (FIGO) invasive cervical cancer. HPV16 DNA was detected in 7 (47%) and HPV18 DNA in 3 (20%) of the cervical cancers. Examination of histologically normal lymph nodes from these 10 patients by nested PCR revealed HPV DNA in 5 (50%) of them; in all cases HPV type in lymph nodes and tumor was the same. Two of these 5 patients had a recurrence (pelvic cavity or lung) and died of cancer, although all 5 had had pelvic radiotherapy after radical hysterectomy and lymphadenectomy. These findings indicate that nested PCR is useful for evaluating early lymph-node involvement retrospectively in HPV-positive cases.     

Efficacy of combination therapy for multiple liver metastases of colorectal cancer

In our hospital, combination therapy, mainly intra-arterial infusion, is performed for multiple liver metastases of colorectal cancer. The median survival time of the combination group (n = 18), the hepatectomy only group (n = 3) and the best supportive care group (n = 7) were 21.7, 12.5 and 6.1 months, respectively. The prognosis of the combination group was significantly better than that in the other groups (p < 0.0001). Univariate analysis against the combination group revealed that serum CEA was a significant prognostic factor (p = 0.0196). Moreover, we divided the combination group into two groups on the basis of serum CEA either below or above 50 ng/ml. The prognosis of the low CEA group (n = 11), whose median survival time was 25.9 months, was significantly better than the high CEA group (n = 7), whose median survival time was 17.8 months (p = 0.0031). It therefore appears that combination therapy may be of no benefit when serum CEA is above 50 ng/ml.     

Case report--efficacy of short-term intraarterial 5-fluorouracil for liver metastasis from colorectal cancer

Hepatectomy has the highest cure rate among the various methods for treating liver metastasis from colorectal cancer. We previously reported that continuous hepatic arterial infusion (HAI) of 5-FU is effective for improving the prognosis of patients with liver metastasis. In this study, we examined the efficacy of short-term continuous HAI of 5-FU for treating liver metastasis from colorectal cancer. A 57-year-old woman with a solitary liver metastasis from rectal cancer was treated by continuous HAI of 5-FU (1,000 mg/day) for 6 days. Her elevated serum CEA level (20.7 ng/ml) then returned to normal. Computed tomography revealed a decrease of 55.6% in the size of the liver tumor. Partial segmentectomy (S7) was subsequently performed. Histological examination of the resected tumor showed marked degeneration, necrosis, fibrosis, and calcification with viable moderately differentiated adenocarcinoma cells. These results suggest that preoperative HAI of 5-FU is safe and worth trying in patients with liver metastasis from colorectal cancer.