Analysis of the origin of the right inferior phrenic artery in 178 patients with hepatocellular carcinoma treated by chemoembolization via the right inferior phrenic artery.

BACKGROUND: No previous report has described the level of the origin of the right inferior phrenic artery (RIPA) based on an analysis of the relationships between the level of the RIPA, the celiac artery (CA), the superior mesenteric artery (SMA), and the right renal artery (RRA) in a series of cases. PURPOSE: To evaluate the origin of the RIPA by retrospectively analyzing angiographic findings in 178 patients with hepatocellular carcinoma (HCC) who underwent transcatheter arterial chemoembolization (TACE) via the RIPA. MATERIAL AND METHODS: In patients treated with intraarterial chemoembolization for HCC, additional superselective chemoembolization of the RIPA branches was necessary in 178 cases. We analyzed the level of the origin of the RIPA in these patients according to the relationships between the level of the origin of the RIPA, the CA, the SMA, and the RRA on angiography. RESULTS: Among the 178 cases, the RIPA arose from 1) the aorta directly in 102 cases (57%), 2) the CA in 53 (30%), 3) the left gastric artery (LGA) in three (2%), 4) the dorsal pancreatic artery (DPA) in one (1%), and 5) the RRA in 19 (11%). The level of the origin of the RIPA that originated directly from the aorta was supraceliac in 56 cases (32%), between the CA and the SMA in 31 (17%), and between the SMA and the RRA in 15 (8%). CONCLUSION: In our study, the RIPA originated from the aorta between the CA and the SMA directly in 17% of cases. When it is difficult to identify the origin of the RIPA, we must keep in mind that the RIPA may originate from the right part of the aorta within the small distance between the SMA and the CA.

A MEK inhibitor (U0126) markedly inhibits direct liver invasion of orthotopically inoculated human gallbladder cancer cells in nude mice.

Primary cancer of the gallbladder is not unusual. Most cases of gallbladder cancer are found at an advanced stage, accompanied by the invasion to the liver, metastases to the lymph nodes and distant organs, and peritoneal dissemination. In this study, we first examined the effect of mitogen-activated protein kinase kinase (MEK) inhibitors on the production of matrix metalloproteinases (MMPs), urokinase-type plasminogen activator (uPA), and tissue inhibitors of metalloproteinases (TIMPs) in a human gallbladder cancer cell line, NOZ cells in vitro. MEK inhibitors (PD98059 and U0126) inhibited the production of MMP-2, MMP-9 and high MW uPA, and upregulated TIMPs (TIMP-1, TIMP-2 and TIMP-3). Subsequently, we examined the effect of U0126 on invasion and metastasis of orthotopically inoculated NOZ cells in nude mice. Direct liver invasion by cancer cells was detected in all of the mice in the control group, but in only one mouse in the U0126-treated group. Most of the primary tumors in the U0126-treated group expanded to the liver, but did not invade into the liver. Vessel invasion in the liver was evident in 4 out of 5 mice in the control group, but in only one mouse in the U0126-treated group. Lymph node metastases and peritoneal dissemination were recognized in all of the mice in both groups. All 5 mice in the U0126-treated group, and 4 out of 5 mice in the vehicle control group, had metastases in the lungs. The present results suggest that a MEK inhibitor, U0126, prolonged the survival of the mice with NOZ tumor by inhibiting direct liver invasion and vessel invasion of the cancer cells via down-regulation of the matrix degrading ability of the cancer cells.

Expression of metastasis suppressor gene (KAI1/CD82) in oral squamous cell carcinoma and its clinico-pathological significance.

In this study, we investigated the expression of the KAI1/CD82 gene in oral squamous cell carcinoma (oral SCC). We studied 43 oral SCC patients. Reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to evaluate expression of this gene, and results were compared to the clinico-pathological findings. Twenty-five specimens (58.1%) were KAI1/CD82-positive, and 18 (41.9%) were negative. There were statistically significant relationships between gene expression and both histological malignancy (P=0.0205) and mode of invasion (P=0.0315). But there were no correlations of expression with tumor status, regional lymph node metastasis, pathological lymph node metastasis or histological differentiation. No significant relationship was observed between patient survival and expression of KAI1/CD82 by tumors. The results of this study suggest that the KAI1/CD82 gene may not be a useful predictor of prognosis, although decreased gene expression may be associated with increased invasive ability of oral SCC.

A new in vivo model for studying invasion and metastasis of esophageal squamous cell carcinoma.

The aim of our current study was to establish an orthotopic inoculation model for studying invasion and metastasis of esophageal squamous cell carcinoma (SCC). Male BALB/c nude mice were used for the experiment. A midline incision was made from the upper to middle abdomen. The abdominal esophagus was carefully exposed. Human esophageal T.Tn SCC cells or human cervical HeLa SCC cells, were injected into the submucosa of the lower esophagus. One of the mice injected with T.Tn cells was sacrificed at 5 weeks, and the remaining five sacrificed at 13 weeks after inoculation. The mice injected with HeLa cells were sacrificed at 3-4 weeks after inoculation. T.Tn cells and HeLa cells formed tumors at the esophagus, but did not metastasize to lymph nodes or lungs. HeLa cells produced peritoneal implants, and directly invaded the stomach and the liver. In the present study, we established a novel orthotopic inoculation model of esophageal SCC. This system is an appropriate and a useful model for studying invasion and metastasis of esophageal SCC, and can also be used as a model for developing therapeutic strategies for esophageal cancer in vivo.

Dihydropyrimidine dehydrogenase mRNA level correlates with the response to 5-fluorouracil-based chemo-immuno-radiation therapy in human oral squamous cell cancer.

The measurement of the intra-tumoral level of thymidylate synthetase (TS), and dihydropyrimidine dehydrogenase (DPD), may be useful in predicting tumor sensitivity to 5-fluorouracil (5-FU). In this study, we examined the mRNA levels of DPD and TS in 28 oral squamous cell carcinomas (SCC), and 22 salivary gland tumors by semi-quantitative reverse transcription polymerase chain reaction. Then we examined the correlation of the responsiveness of the patients with oral SCC to 5-FU with the intra-tumoral levels of DPD and TS mRNA. All specimens were obtained at the biopsy before treatment, and then the patients were treated by oral administration of a 5-FU compound (UFT), the irradiation of cobalt-60 (upto 60 Gy) and injection of an immuno-potentiator (OK-432). Intra-tumoral levels of DPD mRNA in the patients who showed CR (complete response) and PR (partial response) were significantly lower than those in the patients who showed NC (no change). However, intra-tumoral levels of DPD mRNA did not correlate with the local recurrence of the tumor during the observation period after initial treatment with or without surgical resection of the residual tumors. On the other hand, TS mRNA levels in the tumors did not correlate with any clinico-pathological parameters. These observations suggest that intra-tumoral levels of DPD mRNA may predict the tumor response to 5-FU-based chemo-immuno-radiation therapy in the patients with oral SCC.

Design and synthesis of a highly selective EP4-receptor agonist. Part 2: 5-thia and 9beta-haloPG derivatives with improved stability.

Further chemical modification to identify more chemically stabilized EP4-receptor selective agonists was continued. As a result, a further two EP4-receptor selective agonists 5-thiaPGE(1) 2a, 10 and 9beta-chloroPGF(2) analogue 11 were discovered.

Neuraminidase-producing ability of intestinal bacteria isolated from coastal fish.

The neuraminidase-producing ability of bacteria from coastal fish intestines and their environments was determined using a fluorogenic substrate, 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid. Of 836 isolates examined, 758 (90.7%) produced little or no neuraminidase (< 0.0002 U ml-1) while 78 (9.3%) produced >/= 0.0002 U neuraminidase ml-1. Of note, 10% of vibrios from fish intestines could produce neuraminidases, and 9% did it efficiently. This result suggests that the vibrios capable of producing neuraminidases are able to colonize and establish in the intestinal tract of coastal fish to some extent. Additionally, as many as 58% of the Vibrio isolates with high abilities were tentatively identified as Vibrio damsela.

Sustained inward current during pacemaker depolarization in mammalian sinoatrial node cells.

Several time- and voltage-dependent ionic currents have been identified in cardiac pacemaker cells, including Na(+) current, L- and T-type Ca(2+) currents, hyperpolarization-activated cation current, and various types of delayed rectifier K(+) currents. Mathematical models have demonstrated that spontaneous action potentials can be reconstructed by incorporating these currents, but relative contributions of individual currents vary widely between different models. In 1995, the presence of a novel inward current that was activated by depolarization to the potential range of the slow diastolic depolarization in rabbit sinoatrial (SA) node cells was reported. Because the current showed little inactivation during depolarizing pulses, it was called the sustained inward current (I(st)). A similar current is also found in SA node cells of the guinea pig and rat and in subsidiary pacemaker atrioventricular node cells. Recently, single-channel analysis has revealed a nicardipine-sensitive, 13-pS Na(+) current, which is activated by depolarization to the diastolic potential range in guinea pig SA node cells. This channel differs from rapid voltage-gated Na(+) or L-type Ca(2+) channels both in unitary conductance and gating kinetics. Because I(st) was observed only in spontaneously beating SA node cells, ie, it was absent in quiescent cells dissociated from the same SA or atrioventricular node, an important role of I(st) for generation of intrinsic cardiac automaticity was suggested.

The sustained inward current and inward rectifier K+ current in pacemaker cells dissociated from rat sinoatrial node.

1. Myocytes were dissociated from the sinoatrial (SA) node of rat heart using a new enzymatic dissociation technique. Only a small number of isolated SA node myocytes showed regular rhythmic contractions and spontaneous action potentials, and these were used in the present study. 2. The spontaneous action potential was resistant to TTX, and the action potential parameters were similar to those of rabbit and guinea-pig pacemaker cells. Major time- and voltage-dependent currents were the delayed rectifier K+ current IKr, the L-type Ca2+ current ICa,L and the sodium current INa. The hyperpolarization-activated cation current (If) was recorded from approximately 50 % of the cells with hyperpolarization beyond -90 mV. 3. The instantaneous current jump at the onset of a hyperpolarizing pulse showed inward rectification and was largely blocked by Ba2+. This Ba2+-sensitive current corresponded well to the inward rectifier K+ current (IK1), although it was much smaller in amplitude than in the ventricle. 4. A sustained inward current was activated on depolarization from -80 mV to the voltage range of slow diastolic depolarization. The current was blocked by nicardipine, enlarged by isoprenaline and was insensitive to removal of external Ca2+. These characteristics were similar to the sustained inward current, Ist, previously described in the rabbit and guinea-pig SA node cells. 5. The role of Ist was considered by constructing empirical equations, which were applied to the experimental record of the action potential. It is demonstrated that the voltage-dependent activation of Ist constitutes a positive feedback loop with the depolarization of the membrane.

Early establishment of hematopoietic chimerism following allogeneic peripheral blood stem cell transplantation in comparison with allogeneic bone marrow transplantation.

To characterize the process of the establishment of complete chimerism after allogeneic peripheral blood stem cell transplantation (allo-PBSCT), we determined the origin of leukocytes in peripheral blood (PB) obtained from 23 patients in the very early period after allo-PBSCT using amplification of mini- or microsatellite regions of genomic DNA. Donor-specific alleles were amplified from the PB obtained at day 8 post-transplant for 19 allo-PBSCT patients. Among the 19 patients, 12 showed only donor-specific alleles (complete chimerism) while 7 did both donor and host-specific alleles (mixed chimerism). Although donor specific alleles were amplified in 10 of 12 patients who received allogeneic bone marrow transplantation (allo-BMT) similarly to allo-PBSCT, all of these ten showed mixed chimerism. When the chimeric state was examined in PB samples obtained serially at 2-3-day intervals post-transplant, host-specific alleles in allo-PBSCT patients were not detectable in the PB much earlier than those in allo-BMT patients. These findings indicate that the appearance of donor-derived cells associated with the disappearance of host-derived cells in the circulation occurs earlier after allo-PBSCT as compared with allo-BMT, leading to the rapid establishment of complete chimerism.

Cellulitis of the eyelids associated with sinusitis and brain abscess.

Erythema in the orbital area can indicate systemic and life-threatening diseases. We experienced an unusual and serious case of orbital cellulitis that was difficult to distinguish from a case with good prognosis. A 21-year-old man developed an erythema around his eyes. He exhibited no symptoms that would suggest lesions in deep tissues, but his condition turned out to be cellulitis retrogradely metastasized from an odontogenic sinusitis traced to a dental treatment problem. Computed tomography revealed complication of a large abscess in the frontal lobe. Cellulitis of the orbital area requires particular clinical discretion.

Temperature dependence of the inward rectifier K+ channel gating in guinea-pig ventricular cells.

Whole-cell and single-channel currents of the inward rectifier K+ channels from guinea-pig ventricular myocytes were recorded over the range between 5 and 37 degrees C. The conductance for inward currents was decreased by lowering the temperature with a Q10 of 1.28 (whole cell), or 1.41 (single channel) between 20 and 30 degrees C. The open probability of the channel at -100 mV remained high (> 0.9). The distribution of open times was single exponential at all temperatures, confirming a single open state. The entropy change (delta S) for the closing rate of the channel obtained from open-time distribution was -14.0 e.u. (cal/mol/K), and enthalpy change (delta H) was 11.9 kcal/mol. The configuration of closed-time distribution varied markedly by altering the temperature, and three exponentials were necessary to fit the histogram. The slowest component showed higher temperature dependency (delta S = 13.6 e.u. and delta H = 19.0 kcal/mol) than the other two faster components. By assuming a reduced model of C-O at 37 degrees C, the difference in Gibb's free energy (GOC) between the open and closed states was approximately 2 kcal/mol, and the height of the energy barrier for the C-O transition was estimated to be approximately 15 kcal/mol.

[Homozygous beta(+)-thalassemia due to -31 A to G mutation].

A 20-year-old Japanese woman was admitted to our hospital with anemia and mild splenomegaly. Peripheral blood examination revealed Hb 9.4 g/dl, Ht 29.3%, RBC 4.74 x 10(6)/microliters, reticulocytes 2.4%, WBC 5,200/microliters, platelets 24.9 x 10(4)/microliters, MCV 61.7 fl, and MCH 19.9 pg. Poikilocytosis with target cells was recognized on the peripheral blood smear. A bone marrow aspirate revealed erythroid hyperplasia. Serum iron and ferritin were in the normal range. beta-thalassemia was suggested by the increase in HbA2 (6.5%) and HbF (7.5%). Analysis of beta globin DNA by single strand conformation polymorphism (SSCP) and amplification refractory mutation system (ARMS) confirmed a diagnosis of homozygous beta(+)-thalassemia due to -31 A to G mutation. A familial study revealed that her parents were heterozygous for this allele. This is the 8th case of homozygous beta(+)-thalassemia due to -31 A to G mutation in Japan.

Inhibition of thromboxane A2 activity during warm ischemia of the liver.

To determine the role of thromboxane A2 (TxA2) in ischemic damage of the rat liver, we examined the effects of a TxA2 synthetase inhibitor (OKY 046) and a TxA2 receptor antagonist (ONO 3708). Rats were divided into three groups. In group I, a portion of the liver was subjected to 100 min of warm ischemia and the remaining liver resected. In group II, OKY 046 (30 mg/kg, intravenously) was given 5 min before the same procedure. In group III, ONO 3708 (10 mg/kg, intravenous) was given 5 min before ischemia. We then assessed survival, serum biochemistry, extent of histologic necrosis, and the levels of prostaglandin E2 (PGE2), TxB2, and 6-keto-PGF1-alpha. Pretreatment with OKY 046 and ONO 3708 significantly improved survival, decreased the tissue water content, and lowered the levels of serum transaminases and the extent of histological liver necrosis compared with the control group. OKY 046 markedly suppressed the level of TxB2, but not the levels of PGE2 or 6-keto-PGF1-alpha. ONO 3708 did not change the levels of PGE2, TxB2, or 6-keto-PGE1-alpha. In a liver perfusion model, OKY 046 and ONO 3708 did not suppress the uptake of trypan blue in hepatocytes. Our results demonstrate that either a TxA2 synthetase inhibitor or a TxA2 receptor antagonist can protect the liver from an ischemic insult. The effects of these drugs were due to inhibition of TxA2 synthesis and TxA2 blockade at the receptor, without modulating PGI2 or PGE1. Our results in a perfused rat liver model suggest that these drugs work during reperfusion and prevent postischemic tissue edema.

Nucleoside-nucleotide mixture increases peripheral neutrophils in cyclophosphamide-induced neutropenic mice.

The importance of nucleotides and nucleosides in human nutrition has now become an area of intensive research and clinical interest. To determine any potential benefits of administering a nucleoside-nucleotide mixture (NNM) to cyclophosphamide (CPA)-induced neutropenic mice, we randomly assigned 20 BALB/c mice to two groups and fed them a nucleic-acid-free 20%-casein diet for 20 days. The mice were intraperitoneally administered NNM or saline daily from the onset of the experiment. On the 10th day of this treatment, the mice were intraperitoneally injected with CPA. Blood was collected retro-orbitally and differential counts of leucocytes were done on blood smears by counting microscopically on day 0 (before) and 1, 3, 5, 7, and 10 days after CPA injection. By the 7th and 10th days after CPA injection, respectively, the peripheral neutrophil number in the saline group was significantly lower compared with the neutrophil number in the NNM group (p < 0.05). Also in the NNM group, more clusters of matured neutrophils were observed compared with the saline group. These results suggest that NNM may stimulate the proliferation, differentiation, and maturation of neutrophils, and that NNM alone or in combination with other pharmacologic agents may be an important therapeutic agent in patients after chemotherapy, immunosuppressive therapy, and organ transplant.

A mixture of nucleosides and nucleotides increases bone marrow cell and peripheral neutrophil number in mice infected with methicillin-resistant Staphylococcus aureus.

We studied the effects of a mixture of nucleosides and nucleotides on the peripheral neutrophil number and the proliferation of bone marrow cells in mice challenged with methicillin-resistant Staphylococcus aureus. BALB/c mice were fed a nucleotide-free 20% casein diet (control) or this diet supplemented with nucleosides and nucleotides orally (Expt. 1) or intraperitoneally (Expt. 2 and 3). On d 10, the mice were challenged intravenously with methicillin-resistant S. aureus (6.7 x 10(12) colony forming units/L). In Expt. 1 and 2, numbers of total and differential counts of blood leucocytes were counted on d 0 (before), 1, 3 and 5 after the infection. In Expt. 3, 30 min before killing, bromodeoxyuridine (20 mg/kg), an analogue of thymidine, was administered intraperitoneally and its incorporation in the DNA synthetic phase of bone marrow cells was determined at 0 h (before), 3, 6 and 24 h after the infection. Mice fed the supplemented diet had higher (P < 0.05) leucocyte and neutrophil numbers on d 0 compared with the control group. The neutrophil numbers tended to be greater in the supplemented group at 1, 3 and 5 d after the infection. Intraperitoneal administration of nucleosides and nucleotides increased (P < 0.05) neutrophil numbers before and after the infection. Twenty-four h after the infection, incorporation of bromodeoxyuridine into the DNA synthetic phase of bone marrow cells in the group administered nucleosides and nucleotides was higher (P < 0.05) than in the control group. We conclude that, following methicillin-resistant S. aureus injection, intraperitoneal administration of a nucleoside-nucleotide mixture may stimulate bone marrow cell proliferation and increase the peripheral blood neutrophil numbers. Oral administration may elicit weaker effects.

Ganglioside antigen of DU-PAN-2 in a human pancreatic cancer.

DU-PAN-2 reactive gangliosides were isolated from the tumor of a patient with pancreatic cancer (duct cell carcinoma, moderately differentiated adenocarcinoma), having a negative Lewis blood phenotype, and were analyzed by means of TLC-immunostaining, enzyme-linked immunosorbent assay (ELISA), permethylation study, 1H NMR spectroscopy and fast atom bombardment mass spectrometry. The structures of the gangliosides were found to be NeuAc alpha 2-3Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1'Cer, containing normal and hydroxy fatty acids. By TLC-immunostaining and ELISA with chemically synthesized gangliosides, DU-PAN-2 was demonstrated to react strongly with IV3 alpha NeuAc-Lc4Cer, weakly with IV3 alpha NeuAc-nLc4Cer, and moderately with IV6 alpha NeuAc-Lc4Cer and IV6 alpha NeuAc-nLc4Cer. Thus it was concluded that the DU-PAN-2 reactive ganglioside in the tumor is IV3 alpha NeuAc-Lc4Cer and that DU-PAN-2 has a rather broad specificity.