Kidney produces a novel acylated peptide, ghrelin.

Ghrelin is a novel growth hormone-releasing peptide with a unique acylated structure. Here we reveal that prepro-ghrelin gene is expressed in the mouse kidney and glomerulus. We also show by reverse-phase high performance liquid chromatography coupled with radioimmunoassay that the mouse kidney does produce ghrelin. The ghrelin immunoreactivity in the mouse kidney is 6.79+/-0.48 fmol/mg (n=5), which is much more abundant than that in the mouse plasma of 0.339+/-0.029 fmol/microl (n=6). Furthermore, prepro-ghrelin gene is expressed in cultured rat mesangial cells, fibroblast-like NRK-49F cells and mouse podocytes, but not in rat epithelial cell-like NRK-52E cells. Ghrelin receptor gene is also expressed in the rat kidney. These findings demonstrate that the kidney, glomerulus and renal cells express prepro-ghrelin gene and ghrelin is produced locally in the kidney, and suggest the endocrine and/or paracrine roles of ghrelin in the kidney.

Production of a new hybrid anthracycline 4-O-methylepelmycin by heterologous expression of dnrK in epelmycin-producing Streptomyces violaceus.

A new hybrid anthracycline antibiotic was produced by heterologous expression of dnrK encoding carminomycin 4-O-metyltransferase in an epelmycin-producing Streptomyces violaceus. pMK100 was constructed by insertion of Steptomyces peucetius dnrK gene in Steptomyces-expression vector pIJ6021 and introduced to the epelmycin producer. The transformant produced a hybrid anthracycline antibiotic together with host epelmycins when cultured in antibiotic production medium in the presence of thiostrepton. The hybrid anthracycline was determined to be 7-O-L-rhodosaminyl-4-O-methyl-epsilon-rhodomycinone (4-O-methylepelmycin D). However, the attempts on production of hybrid 4-O-methylaclarubicin and 4-O-methyl-1-deoxyobelmycin by the transformants of aclarubicin and 1-deoxyobelmycin producers with pMK 100 were unsuccessful.

Effects of bacterial glyceroglycolipid M874B on growth and TPA-induced differentiation of HL60 cells.

Bacterial monogalactosyldiacylglycerol M874B (MGDAG), which protects against oxygen radicals, was found to increase the growth of the human promyelocytic leukemia cell HL60 when added to the cell culture, but suppresses the 12-O-tetradecanoyl phorbol-13-acetate-induced differentiation. Analogous MGDAG, S365B had weak, but similar effects. These activities were not observed with analogous plant glyceroglycolipids and diacylglycerol.

Effects of olprinone on systemic and cerebral circulation in patients with subarachnoid hemorrhage.

Olprinone is a promising new drug used for treating cerebral vasospasm. To clarify the effects of olprinone on systemic and cerebral circulation in patients with subarachnoid hemorrhage, hemodynamic and oxygenation parameters were evaluated as 12 such patients underwent surgery. After aneurysm clipping and confirmation of hemostasis, olprinone was administrated at a dose of 10 microg/kg over 5 minutes followed by 0.2 microg/kg/min for 25 minutes. Variables before and after administration were compared by paired t tests. Heart rate and cardiac index increased while no significant changes occurred in oxygen saturation of mixed venous blood, or oxygen extraction ratio. Cortical blood flow increased and cerebral vascular resistance decreased significantly, but oxygen saturation in the jugular bulb, arterio-jugular difference of oxygen content, and lactate oxygen index did not change significantly. In conclusion, olprinone increased cardiac output and cortical blood flow in patients with subarachnoid hemorrhage, but the balance between oxygen supply and consumption systemically and in the brain did not change. This observation suggests the possibility that olprinone increases cerebral metabolism.

Ghrelin strongly stimulates growth hormone release in humans.

Ghrelin is a recently identified endogenous ligand for the GH secretagogue receptor and is involved in a novel system for regulating GH release. However, little is known about its GH-releasing activity and other endocrine effects in humans. To address this issue, we studied the GH, ACTH, cortisol, PRL, LH, FSH, and TSH responses to synthetic human ghrelin. In four normal male adults (28-37 yr), iv ghrelin administration released GH in a dose-dependent manner and 0.2, 1.0, and 5.0 microg/kg ghrelin produced 43.3 +/- 6.0, 81.5 +/- 12.7, and 107.0 +/- 10.7 ng/mL of the GH peak values at 30 min, respectively. ACTH, cortisol, and PRL levels were also elevated after ghrelin injection, while the lowest dose (0.2 microg/kg) resulted in only minimum peak values of these hormones (22.8 +/- 3.0 pg/mL, 9.4 +/- 1.9 microg/dL, and 4.6 +/- 0.6 ng/mL, respectively). There were no significant changes in LH, FSH, or TSH levels. This is the first study showing evidence that ghrelin strongly stimulates GH release in humans.

[Anatomy of intrahepatic portal branches visualized by three-dimensional imaging analysis of CT arterial portography].

The purpose of this study was to clarify variations in intrahepatic portal branches by means of CT imaging procedures. The subjects were 73 patients, 59 men and 14 women, who ranged in age from 41 to 76 years, with a mean of 63 years. The procedures were as follows. The entire liver was scanned using helical CT during the portal and hepatic venous phases, and 3D images of the portal vein were reconstructed with the volume-rendering technique and the region-growing method. The CT unit was a HITACHI W2000, and the imaging analyzer a Sun Ultra 1. We found that the branching patterns of both the anterior (P5 and P8) and posterior segmental branches (P6 and P7) of the right lobe of the liver could be classified into four types. The caudate branch (P1) and left lateral segmental branches (P2 and P3) were classified into three types, and the interior segmental branch of the left lobe (P4) was classified into two types. The frequency of each pattern was also revealed. These branching types and their frequencies were generally the same as those described in previous reports. Thus, the portal anatomy visualized by these methods indicates that they could be very useful for preoperative examinations or IVR.

Finger bougie method compared with pyloroplasty in the gastric replacement of the esophagus.

To elucidate the necessity of pyloroplasty for the gastric tube through the posterior mediastinum in esophageal surgery, gastric emptying and duodenogastric reflux (DGR) were evaluated in 16 cases undergoing an anterior pylorectomy (group P) and in 16 cases treated by the finger bougie method (group F). First, the obstruction and reflux symptoms were examined based on a patient questionnaire using a brief scoring system. The median value of the symptom score showed the patients in P to have more symptoms than those in F; however, the difference was not significant (8.0 vs 6.0). Secondly, the swallowed Tc O4- (85MBq) was counted using a gamma camera at three sites on the sternal bone in the upright position based on a gastric transit scintigram. Both the descending time of the RI peak and the clearance rates were similar between the two groups. Thirdly, intragastric 24-h pH monitoring was carried out. Antimony pH sensors were anchored 5 and 15cm below the esophagogastrostomy. We could not find any difference between the two groups in both the %time pH > 4 and %time pH > 7. These findings thus revealed no big difference between groups P and F. The finger bougie method to drain the vagotomized posterior mediastinal stomach was found to achieve results similar to conventional pyloroplasty, while it was also simpler and safer.

[Five patients with intrapulmonary lymph nodes indistinguishable from small peripheral carcinoma of the lung].

We encountered 5 patients with subpleural intrapulmonary lymph nodes. In all 5 patients, computed tomographic (CT) scans demonstrated subpleural small nodules in the middle or lower lung lobes. Because lung cancer could not be ruled out, biopsies by video-assisted thoracic surgery (VATS) were performed, yielding a diagnosis of intrapulmonary lymph nodes in all 5 cases. Two of the patients were women and nonsmokers. Sharply defined borders and subpleural locations were the characteristic CT findings for the intrapulmonary lymph nodes. However the borders of some of the nodules were irregular. Small peripheral lung cancer is also sometimes characterized by sharply defined borders and subpleural locations. For this reason, it is difficult to differentiate between intrapulmonary lymph nodes and small peripheral lung cancer on the basis of CT findings alone. VATS should be readily employed for diagnostic purposes in such cases.

[Arterial infusion of SMANCS-Lipiodol for advanced hepatocellular carcinoma].

Twenty-four patients were treated with arterial infusion of SMANCS dissolved in Lipiodol. Twenty of these patients had HCC with the main trunks of portal vein occluded by tumor, and four patients had severe cirrhosis and multiple HCC. The actual dose of SMANCS administered each patient ranged from 4 to 6 mg. Side effects occurred in 50%. Severe side effects such as shock and shivering-chilliness were observed in 18%. The differences between the values of hepatic functional serum indexes obtained before and after treatment with SMANCS were small and transient. With regard to the therapeutic response of the arterial infusion of SMANCS, the mean survival time was approximately 2.8 months. It was suggested that the more effective administration of SMANCS was combination of the arterial infusion of SMANCS-Lipiodol with TAE at the level of the right hepatic artery of left hepatic artery for multiple HCC.

Effects of nucleotides on cyanide-resistant respiratory activity in mitochondria isolated from antimycin A-treated yeast Hansenula anomala.

Mitochondria were isolated from Hansenula anomala induced for cyanide-resistant respiration by antimycin A-treatment. Cyanide-resistant respiratory activity in isolated mitochondria was stimulated by AMP, ADP, dAMP, and GMP, but not by ATP, adenosine, cAMP, 2'(3')-AMP, CMP, and UMP. Effects of nucleotides were also observed on cyanide-resistant and salicylhydroxamate-sensitive decylubiquinol oxidase activity. Carbonylcyanide m-chlorophenylhydrazone, oligomycin, and carboxyatractyloside did not affect activation of decylubiquinol oxidase activity by AMP. It is suggested that purine nucleoside 5'-monophosphate or diphosphate stimulates alternative oxidase activity from the outer surface of the mitochondrial inner membrane with a mechanism different from respiratory control. Alternative oxidase activity might be controlled by adenine nucleotides posttranscriptionally in fungi.

Enhancing effect of thoraco-laparotomy on liver metastasis and the role played by active oxygens in its mechanism.

The enhancing effect on liver metastasis produced by the excessive surgical stress of thoraco-laparotomy (TL), and its regulation with a radical scavenger, were studied in 10-week-old Donryu rats. The rats were divided into three groups: those given thoraco-laparotomy for 1 h (the TL group); those given laparotomy alone for 1 h (the L group); those given a short laparotomy (the C group). The effects of treatment with 5 mg/kg of EPC-K1 was assessed in the TL group. A rat hepatocellular carcinoma cell line AH 60C (5 x 10(5) cells) was administered into the portal vein under general anesthesia. The number of metastatic liver nodules was counted 3 weeks later, and the lipid peroxide (LPO) levels of the liver and serum were measured by the TBA method on postoperative days (PODs) 1, 2, and 3. The number of metastatic liver nodules was 40.6 +/- 29.7, 15.0 +/- 15.8, and 13.7 +/- 9.4 in the TL, L, and C groups, respectively. When EPC-K1 was administered to the TL group, the LPO level on POD 1 decreased from 49.8 +/- 25.8 to 18.9 +/- 7.9 nM/g, and the number of metastatic liver nodules decreased from 27.2 +/- 30.0 to 8.9 +/- 12.7 in parallel. The findings of this study suggested that the excessive surgical stress produced by thoraco-laparotomy enhanced liver metastasis in parallel with an increase in LPO levels; however, the radical scavenger EPC-K1 could aid in reversing this effect.

[Usefulness of subcutaneous implantable port with new one-way valve].

Because of the development of the subcutaneous implantable port, intra-arterial infusion chemotherapy for malignant tumor has been performed in safety and it is now possible to choose a more effective method of drug administration. Many problems remain, however. In this study, we investigated the performance of subcutaneous implantable port with a new one-way valve with safety, septum intensity and a back stream of blood. This new port was very safe, with good septum intensity and water backflow. In the performance test with a back stream of water into the catheter, water did not back up into the catheter connecting with the port with the one-way valve, but did so without the one-way valve after releasing a load of 2,000 g from the septum. This suggests that the catheter obstruction is decreased by adding a one-way valve to the port.

[Repair of the gastric tube-right main bronchus fistula after operation for esophageal cancer--treatment by transposition of pedicled pectoralis major muscle flap].

We have experienced a case who showed the gastric tube-right main bronchus fistula. A 51-year-old male complained cough and vomiting suddenly. He underwent esophagectomy and radio-chemotherapy for advanced esophageal cancer 19 months ago. Chest X-ray showed severe pneumonia, and gastroscopy, bronchoscopy and CT scan showed the fistula between the whole stomach esophageal substitute and right main bronchus. After recovery from the pneumonia with the treatment by continuous suction through the naso-gastric tube, operation was performed. The fistula was repaired with transposition of a pedicled pectralis major muscle successfully. After the operation, respiration was performed independently with two ventilators for right and left lung to avoid increasing air way pressure. His postoperative course was uneventful, and he discharged on the 66th postoperative day. The cause of the fistula was considered to be a peptic ulcer due to residual secretion of gastric acid.

New betaclamycin and aclarubicin analogs obtained by prolonged microbial conversion with an aclarubicin-negative mutant.

Microbial conversion of beta-rhodomycinone and aklavinone using an aclarubicin-negative Streptomyces galilaeus mutant afforded new anthracycline antibiotics CG21-C and CG1-C which had a rednosyl-2-deoxyfucosyl-rhodosaminyl trisaccharide residue at C-7 of each added aglycone. They were produced only when a prolonged conversion culture took place. Because the usual conversion products containing a cinerulosyl-2-deoxyfucosyl-rhodosaminyl residue were at first accumulated and then decreased during further cultivation, it was evident that they occurred by the modification of terminal cinerulose. The isolation, purification, and structural determination are described, and cytotoxicity in vitro against cultured L1210 cells and the formation mechanism are discussed.

New anthracycline antibiotics 10-epi-oxaunomycin and 10-epi-11-deoxyoxaunomycin.

Two new anthracycline antibiotics, 10-epi-oxaunomycin and 10-epi-11-deoxyoxaunomycin, were photochemically obtained from anthracycline metabolites D788-1 (10-carboxy-13-deoxocarminomy-cin) and D788-3 (10-carboxy-11-deoxy-13-deoxocarminomycin) and were examined for their growth inhibitory activities on cultured L1210 leukemic cells. Effects of the S configuration of C-10 and a hydroxyl group at C-11 on the bioactivity are discussed in comparison with oxaunomycin and 11-deoxyoxaunomycin.

Preservation of the lower esophageal sphincter during total gastrectomy for gastric cancer to prevent postoperative reflux esophagitis.

The lower esophageal sphincter (LES) is usually removed during total gastrectomy to successfully perform a curative operation. In this study, the preservation of the LES in curative total gastrectomy was attempted to reduce the reflux. An experimental study using dogs has revealed that the high-pressure zone of the LES can be preserved by making a resection at the gastroesophageal junction, which thus helps to protect the reflux. A previous clinicopathological study revealed that the LES can be preserved without any fear of recurrence at the resection site, if the tumor is located more than 2.0 cm and 3.0 cm from the gastroesophageal junction to the oral margin in node-negative and -positive cases, respectively. Clinically, 8 patients underwent an LES-preserving total gastrectomy [LES(+) gastrectomy] while 19 had an LES(-) gastrectomy in the same period. Of the five LES(+) cases examined, all showed a high pressure zone, whereas none of the four LES(-) cases examined showed such a high-pressure zone after the operation. Endoscopic examination showed that only one of the seven LES(+) cases but six of nine LES(-) cases revealed esophagitis.

New rhodomycin analogs, SS-288A and SS-288B, produced by a Streptomyces violaceus A262 mutant.

Two new rhodomycin metabolites, SS-288A and SS-288B, were specifically produced by a blocked mutant obtained from Streptomyces violaceus A262 and were respectively identified as 7,10-di(O-rhodosaminyl-deoxyfucosyl-deoxyfucosyl)-beta -rhodomycinone and -beta-isorhodomycinone.

Clinical usefulness of an ATP heat sensitivity test using endoscopic biopsy materials from esophageal cancer patients.

The usefulness of a heat sensitivity test which involved performing an ATP assay on endoscopic biopsy materials for predicting the clinical response to hyperthermia was investigated in esophageal cancer patients. Following in vitro heat treatment of FM3A tumor cells, the heat sensitivity detected by ATPA was significantly correlated with that in the colony-forming assay, and the percent inhibition of the ATP level in the tumor cells was correlated with in vivo tumor growth. The heat sensitivity of the biopsy materials evaluated by ATPA correlated well with that of the resected specimens in 18 esophageal cancer patients, while the clinical response to thermal therapy was well predicted by the heat sensitivity of the biopsy materials evaluated by ATPA in seven of the patients. These results indicate that the heat sensitivity test conducted by performing an ATPA on endoscopic biopsy materials could be a useful indicator for predicting the clinical response to thermal therapy.