A novel heterozygous missense mutation in the vasopressin moiety is identified in a Japanese person with neurohypophyseal diabetes insipidus.

The autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI) is caused by diverse mutations in one allele of the gene that encodes the arginine vasopressin (AVP) precursor protein, AVP-neurophysin II (AVP-NP II). Most of the mutations identified so far are located in either the signal peptide or NP II moiety. Two recently published mutations in the AVP gene identified in kindreds with adFNDI predict a substitution of histidine for tyrosine at position 2 and a deletion of phenylalanine at position 3 in AVP. They are unique among adFNDI mutations in that they are the only adFNDI mutations that affect amino acid residues in the AVP moiety of the pro-hormone. Here, we report a novel heterozygous missense mutation in the AVP moiety of the AVP-NP II gene in a Japanese person with neurohypophyseal diabetes insipidus (DI). This mutation occurs at position 2 in AVP and predicts a substitution of serine for tyrosine (Y21S). It is expected to interfere with normal binding of AVP with NP II, and thus result in misfolding of the precursor proteins. The data of this study support the notion that mutations affecting the AVP moiety can result in the initiation of the pathological processes.

A unique phosphatidylinositol bearing a novel branched-chain fatty acid from Rhodococcus equi binds to influenza virus hemagglutinin and inhibits the infection of cells.

From the aquatic bacterium Rhodococcus equi strain S(420), we isolated a substance that strongly binds to influenza viruses. Structural analyses revealed that it is a unique type of phosphatidylinositol (PtdIns) bearing a branched-chain fatty acid (14-methyloctadecanoic acid). In a TLC/virus-binding immunostaining assay, this PtdIns bound to all subtypes of hemagglutinin (HA) of influenza A viruses tested, isolated from humans, ducks and swine, and also to human influenza B viruses. Furthermore, the PtdIns significantly prevented the infection of MDCK cells by influenza viruses, and also inhibited the virus-mediated hemagglutination and low pH-induced hemolysis of human erythrocytes, which represents the fusogenic activities of the viral HA. We also used purified hemagglutinin instead of virions to examine the interaction between viral HA and PtdIns, showing that the PtdIns binds to hemagglutinin. These findings indicate that the inhibitory mechanism of PtdIns on the influenza virus infection may be through its binding to viral HA spikes and host cell endosomal/lysosomal membranes, which are mediated by the function of viral HA.

[A case of pneumonia, septic pulmonary embolisms with cavities, bilateral large lung abscesses, and multiple liver abscesses caused by Klebsiella pneumoniae].

A 64-year-old man suffering from diabetic hyperosmolar non-ketotic coma was admitted for acute lung abscess in the left apical lung field. Sputum culture and blood culture showed a heavy growth of Klebsiella pneumoniae (K. pneumoniae). He was suffering from sepsis, septic pulmonary embolisms with cavities, bilateral pulmonary consolidations, and multiple liver abscesses. Gradually, the bilateral lung consolidations resolved and areas of consolidation were noted to undergo extensive cavitation bilaterally. Cavitation and abscess formation are frequent complications of K. pneumoniae. Generally, large bilateral lung abscesses caused by K. pneumoniae have a poor prognosis. Cavity nodules are often present in septic pulmonary embolisms. We report a very rare case in a patient with three types of cavities with differing mechanisms. The first was an acute lung abscess, the second, septic pulmonary embolisms with cavities, and the third, large bilateral lung cavities noted in the course of resolving consolidations.

Analysis of structural components and molecular construction of soybean soluble polysaccharides by stepwise enzymatic degradation.

Soybean soluble polysaccharides (SSPS) extracted from soybean cotyledons have a pectin-like structure. The core polysaccharides after treatments with four kinds of hemicellulases and a pectinase contained approximately equal numbers of L-rhamnose and D-galacturonate residues, suggesting the presence of the rhamnogalacturonan (RG) I structure consisting of the diglycosyl repeating unit, -4)-alpha-D-GalpA-(1-->2)-alpha-L-Rhap-(1-. The lengths of RG chains were calculated as approximately 15, 28, and 100 diglycosyl repeats. The RG components linked to each other by intervention of galacturonan (GN) chains, constituting the backbone of SSPS. All arabinose residues, which constitute 21% of total SSPS sugars, were found to be in side chains from RG regions, and this was also true for galactose residues, which constitute 50% of total sugars. Of arabinose residues, 94% are present as alpha-1,3- or alpha-1,5-arabinans, and 89% of galactose residues were present as beta-1,4-galactans. Galactan chains are modified with arabinose, xylose, fucose, and glucose at the sites close to the RG regions.

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.

A new type of glycoglycerolipids from Corynebacterium aquaticum.

A new type of glycoglycerolipids, S361A and S365A, were obtained from Corynebacterium aquaticum strains, S361 and S365, newly isolated from soils, and were identified as (2R)-1-[alpha-glucopyranosyl-(1alpha-3)-(6O-acyl-alpha-manno pyranosyl)]-3-O-acylglycerol and (2R)-1-[alpha-mannopyranosyl-(1alpha-3)-(6-O-acyl-alpha-mannopyran osyl)]-3-O-acylglycerol, respectively. S365A was identical to a novel glycoglycerolipid recently isolated from some bacteria, but S361A was a new analog having a glucosylmannosyl in place of the dimannosyl group. Our results indicate that this sn-2 lysotype of glyceroglycolipids may be widely distributed in bacteria.

Influenza A virus-binding activity of glycoglycerolipids of aquatic bacteria.

As the aqueous sphere has been proposed to be an important source medium for the virus infection of land animals, the glycolipids of some aquatic organisms were examined for human influenza A virus-binding activity. Active compounds were not found among the eight echinoderm gangliosides, but two active non-sialylated glycoglycerolipids were isolated from an aquatic bacterium, Corynebacterium aquaticum. The structural formula of one of them, H632A, was elucidated to be 1-14-methyl-hexadecanoyl-3-alpha-D-galactopyranosyl-(1-->3)-6-(12-met hyl-tetradecanoyl)-1-alpha-D-mannopyranosyl]-sn-glycerol. The latter together with reported one elsewhere, S365A, 1-14-methyl-hexadecanoyl-3-[alpha-D-mannopyranosyl-(1-->3)-6-(12-meth yl-tetradecanoyl)-1-alpha-D-mannopyranosyl]-sn-glycerol, apparently bound to three human influenza viruses, A/PR/8/34 (H1N1), A/Aichi/2/68 (H3N2), and A/Memphis/1/71 (H3N2), exhibiting 7-12% (H632A) and 10-22% (S365A) of the activities of the control substances (Neu5Acalpha2-3-paragloboside and Neu5Acalpha2-6- paragloboside). Additionally, these glycolipids were assumed to have virus-neutralizing activities for the following two reasons: (i) The hemagglutination and hemolysis activities of the viruses were inhibited by the glycolipid. (ii) The leakage of a cytosolic enzyme (lactate dehydrogenase) from Madin-Darby canine kidney cells on virus infection was prevented by the glycolipids to nearly the same extent as by fetuin. This is the first evidence of the binding- and neutralizing-abilities of native glycoglycerolipids as to influenza viruses.

Glyceroglycolipids preventing tert-butylhydroperoxide-induced cell death from Microbacterium sp. and Corynebacterium aquaticum strains.

Galactosyl diacylglycerols M874B and S365B obtained from the recently isolated bacteria identified as Microbacterium sp. M874 and Corynebacterium aquaticum S365 were found to prevent oxidative cell death induced by tert-butylhydroperoxide. Their structures were determined to be 1,2-di-O-(12-methyltetradecanoyl)-3-O-beta-D-galactopyranosyl-sn-glycerol and 1-O-(14-methylhexadecanoyl)-2-O-(12-methyltetradecanoyl)-3-O-beta-D-galactopyranosyl-sn-glycerol, respectively.

Protective effects of bacterial glyceroglycolipid M874B against cell death caused by exposure to heat and hydrogen peroxide.

It was revealed by bioassay using sodA and katA mutants of Bacillus subtilis that the bacterial monogalactosyldiacylglycerol M874B, previously characterized as an alkyl peroxyl radical scavenger, was also capable of protecting cells from death caused by heating and exogenous H2O2. Chemical assays using the Fenton reaction and xanthine-xanthine oxidase revealed that M874B could quench hydroxyl radicals but not superoxide anions. Wheat monogalactosyldiacylglycerol, but neither digalactosyldiacylglycerol nor synthetic diacylglycerol, also had the same activities as those of M874B, although it was less efficient than M874B. These results suggest that monogalactosyldiacylglycerols such as M874B are a new type of oxygen radical scavengers capable of quenching some reactive oxygen species.

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.

The cadherin-like protein is essential to specificity determination and cytotoxic action of the Bacillus thuringiensis insecticidal CryIAa toxin.

The Bacillus thuringiensis CryIAa toxin binds a cadherin-like protein (BtR175) on the brush-border membranes of the Bombyx mori midgut columnar cells, which are the targets. By introducing the BtR175 gene with a baculovirus, Spodoptera frugiperda Sf9 cells expressed BtR175 protein on the cell membrane and became susceptible to the CryIAa toxin. The toxin bound the cadherin repeat adjacent to the membrane and made a pore that passed inorganic ions, causing the cell to swell and burst. This was not observed with a BtR175 variant lacking the toxin-binding site. This in vitro experiment mimicked the specific insecticidal action of the toxin in vivo well.