Evaluation of immune system function in neonatal pigs born vaginally or by Cesarean section.

Full term crossbred sows were selected to study the interaction of the immune system, hypothalamus-pituitary-adrenal axis, and growth in pigs born by Cesarean section (c-section; n=4 sows) or vaginal birth (n=4 sows). Gestation length and birth weight did not differ between vaginal birth and c-section pigs (P=0.34 and 0.

Enzymatic characterization of 5-methylthioribulose-1-phosphate dehydratase of the methionine salvage pathway in Bacillus subtilis.

5-Methylthioribulose-1-phosphate (MTRu-1-P) dehydratase catalyzes the reaction from MTRu-1-P to 2,3-diketo-5-methylthiopentyl-1-phosphate (DK-MTP-1-P) in the methionine salvage pathway in Bacillus subtilis. The properties of this enzyme remain to be determined. We characterized these properties using a recombinant protein.

A long-lived M-like state of phoborhodopsin that mimics the active state.

Pharaonis phoborhodopsin (ppR, also called pharaonis sensory rhodopsin II) is a seven transmembrane helical retinal protein. ppR forms a signaling complex with pharaonis Halobacterial transducer II (pHtrII) in the membrane that transmits a light signal to the sensory system in the cytoplasm. The M-state during the photocycle of ppR (lambda(max) = 386 nm) is one of the active (signaling) intermediates.

Preparation and cytotoxic activity of poly(ethylene glycol)-modified poly(amidoamine) dendrimers bearing adriamycin.

We have developed poly(amidoamine) (PAMAM) dendrimers that have poly(ethylene glycol) (PEG) grafts at all dendrimer chain ends. To obtain PEG-modified dendrimers with sites for conjugation of anticancer drugs for this study, we prepared PAMAM G4 dendrimers that have a glutamic acid (Glu) residue at every chain end of dendrimer; PEG chains were attached to amino groups of Glu residues. We then combined the anticancer drug adriamycin to side chains of the Glu residues using an amide bond, [PEG-Glu(ADR)-G4], or hydrazone bond, [PEG-Glu(NHN-ADR)-G4].

Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension.

Reactive oxygen species (ROS) produced by NADPH oxidase play critical roles in various cellular activities, including plant innate immunity response. In contrast with the large multiprotein NADPH oxidase complex of phagocytes, in plants, only the homologs of the catalytic subunit gp91phox and the cytosolic regulator small GTPase Rac are found. Plant homologs of the gp91phox subunit are known as Rboh (for respiratory burst oxidase homolog).

Generation of highly potent nonviral gene vectors by complexation of lipoplexes and transferrin-bearing fusogenic polymer-modified liposomes in aqueous glucose solution.

We reported previously that complexation of lipoplexes containing 3,5-dipentadecyloxybenzamidine (TRX-20) and transferrin-bearing succinylated poly(glycidol) (SucPG)-modified liposome, which becomes fusogenic under weakly acidic conditions, might produce gene carriers with high transfection activity. For the present study, we prepared the lipoplex-SucPG-modified liposome complexes by mixing them either in phosphate-buffered saline or in an aqueous 5% glucose solution. The complexes prepared in phosphate-buffered saline have large particles of more than 800 nm, whereas the complexes prepared in the glucose solution were remarkably small: 200-300 nm.

Structural and physicochemical features on the metal-mediated base pairs.

The chemical structure of the mercury-mediated T-T pair (T-Hg(I)I-T) was determined with (15)N NMR spectroscopy. In order to determine the chemical structure of the T-Hg(I)I-T pair, (15)N-(15)N J-coupling across a metal center (2JNN) was employed. Notably, this is the first observation of (2)J(NN) in a biological macromolecule (DNA duplex).

Structural analysis of the phototactic transducer protein HtrII linker region from Natronomonas pharaonis.

Halobacterial pharaonis phoborhodopsin [ppR, also called Natronomonas pharaonis sensory rhodopsin II (NpSRII)] is a phototaxis protein which transmits a light signal to the cytoplasm through its transducer protein (pHtrII). pHtrII, a two-transmembrane protein that interacts with ppR, belongs to the group of methyl-accepting chemotaxis proteins (MCPs). Several mutation studies have indicated that the linker region connecting the transmembrane and methylation regions is necessary for signal transduction.

Cilostazol inhibits monocytic cell adhesion to vascular endothelium via upregulation of cAMP.

Aim: Cilostazol is clinically used as an inhibitor of platelet aggregation. Although several reports have demonstrated its anti-inflammatory effect, its effect on monocytes and their adhesive interaction to vascular endothelium remains unclear. We thus examined the potential role of cilostazol towards monocyte endothelial interaction under physiological flow conditions.

The characteristics of relapse in adult-onset minimal-change nephrotic syndrome.

Background: Although minimal-change nephrotic syndrome (MCNS) is highly steroid-responsive, the frequency of relapses in some patients is high, necessitating the administration of repeated courses of prednisolone in high doses. It is, therefore, necessary to identify factors that can predict this increased risk of relapse in some patients in order to establish useful treatment methods to reduce the risk.

Methods: To clarify the factors that might increase the risk of relapses, the data of 82 Japanese adult patients with MCNS receiving treatment at our department were analyzed retrospectively.

Angiotensin-converting enzyme inhibitor attenuates monocyte adhesion to vascular endothelium through modulation of intracellular zinc.

To elucidate an anti-inflammatory role of angiotensin-converting enzyme inhibitors (ACEIs) in cardiovascular disease, we studied the effect of ACEIs in monocyte adhesion to endothelial cells and underlying molecular mechanisms. Treatment of human monocytic THP-1 cells with monocyte chemoattractant protein-1 (MCP-1; 100 ng/ml; 10 min) significantly increased their adhesion to human umbilical vein endothelial cells (HUVECs) under flow condition (P < 0.001).

Enzymatic characterization of 5-methylthioribose 1-phosphate isomerase from Bacillus subtilis.

The product of the mtnA gene of Bacillus subtilis catalyzes the isomerization of 5-methylthioribose 1-phosphate (MTR-1-P) to 5-methylthioribulose 1-phosphate (MTRu-1-P). The catalysis of MtnA is a novel isomerization of an aldose phosphate harboring a phosphate group on the hemiacetal group. This enzyme is distributed widely among bacteria through higher eukaryotes.

Temporal relationships between plasma cortisol, corticosteroid-binding globulin (CBG), and the free cortisol index (FCI) in pigs in response to adrenal stimulation or suppression.

The objective of this study was to document changes in plasma concentrations of total cortisol, porcine corticosteroid-binding globulin (pCBG), and the free cortisol index (FCI) in pigs over a 6-h period in response to adrenal stimulation or suppression. Twenty-four 8-week old pigs allotted in equal numbers were administered ACTH, dexamethasone or saline, and blood samples were collected every 15 min via an indwelling jugular catheter for 1 h prior to and 5 h following treatment. Total plasma cortisol increased in ACTH-treated pigs and decreased in dexamethasone-treated pigs within 0.

Synthesis of poly(amidoamine) dendron-bearing lipids with poly(ethylene glycol) grafts and their use for stabilization of nonviral gene vectors.

Recently, we developed a new type of cationic lipid that consists of an amine-terminated poly(amidoamine) dendron and two long alkyl groups. These dendron-bearing lipids achieved efficient gene transfection of cells through synergetic action of the proton sponge effect and membrane fusion in combination with fusogenic lipid dioleoylphosphatidylethanolamine. Using those dendron-bearing lipids as a base material, we developed in this study a functional component of gene vectors that stabilizes lipoplexes by multiple PEG chains and promotes gene transfection through the proton sponge effect.

Effects of weaning and weaning weight on neuroendocrine regulators of feed intake in pigs.

A depression in feed intake and growth often occurs in the weaned pig. Spray-dried plasma is often added to nursery diets in an attempt to stimulate feed intake during this lag. The current study evaluated gene expression of appetite regulators in hypothalamus and adipose tissue 4 d after weaning.

Alkyl chain moieties of polyamidoamine dendron-bearing lipids influence their function as a nonviral gene vector.

We recently developed a novel family of cationic lipids consisting of a polyamidoamine (PAMAM) dendron and two dodecyl chains. Their transfection activity increases with increasing generation of the dendron moiety [Takahashi et al. (2003) Bioconjugate Chem.

Preparation of poly(ethylene glycol)-modified poly(amido amine) dendrimers encapsulating gold nanoparticles and their heat-generating ability.

Loading of HAuCl4 in poly(amido amine) G4 dendrimers having poly(ethylene glycol) (PEG) grafts at all chain ends and subsequent reduction with NaBH4 yielded PEG-modified dendrimers encapsulating gold nanoparticles (Au NPs) of ca. 2 nm diameter. The Au NPs held in the dendrimers were stable in aqueous solutions and dissolved readily, even after freeze-drying.

Preparation of poly(ethylene glycol)-attached dendrimers encapsulating photosensitizers for application to photodynamic therapy.

Photodynamic therapy (PDT) is a noninvasive treatment of some diseases including cancer. We have developed poly(ethylene glycol) (PEG)-attached dendrimers as a drug-carrier candidate. In this study, we prepared nanocapsules of photosensitizers using PEG-attached dendrimers for application to PDT.

Effect of polycarboxylate blocks on the amidase activity of trypsin through complexation with PEG/polycarboxylate block ionomers.

The amidase reaction of trypsin, which is a member of the serine proteinase family, is accelerated by its complexation with block ionomers containing a polycarboxylate block, such as PEG-PAA, PEG-PGA, or PEG-PMA. PEG-PAA and PEG-PGA had similar effects, causing an increase in the k(cat) value and a shift in the pH profile to a lower pH region. On the other hand, PEG-PMA showed not only an increase in the k(cat) value, but also a decrease in the activation energy; however, there was no shift in the pH dependence of the initial reaction rate.

Studies of DNA recognition mechanism of transcription factor IRF-4.

Transcription factor IFN regulatory factor-4 (IRF-4) prefers a DNA sequence including CCGAAA, though the consensus DNA-binding sequence of the IRF family proteins is NNGAAA, and the crystal structure of PU.1/IRF-4/DNA (GTGAAA) ternary complex indicates the NN region of DNA does not interact with IRF-4 directly. This suggests that there is an indirect DNA recognition mechanism in IRF-4.

Structural analyses on the mercuryII-mediated T-T base pair.

Recently, it was reported that T-T mismatches can specifically recognize Hg(II), and form T-Hg(II)-T pairs. In order to understand the structure and properties of the T-Hg(II)-T pair, we measured NMR spectra for a DNA duplex, d(CGCGTTGTCC) x d(GGACTTCGCG), with two successive T-T mismatches (Hg(II)-binding sites) in the middle of the duplex. We identified imino proton resonances of the T-T mismatches in mercury-free duplex, and performed titration experiments with Hg(II) by using 1-dimensional (1D) (1)H NMR spectra.

NMR structural analysis of the G.G mismatch DNA complexed with naphthyridine-dimer.

Naphthyridine-dimer (ND) specifically recognizes G.G mismatch DNA (Figure 1). However, its detailed recognition mechanism is not clear.

Spectroscopic analyses of DNA duplexes in the presence of mercury ions.

The DNA duplex with tandem T x T mismatches in the presence of Hg(II) has been studied by NMR spectroscopy. For this study, we synthesized decamer duplex with two successive T x T mismatches (TT10). From two-dimensional (2D) 1H-1H NOESY spectrum of TT10 in complex with Hg(II), we were able to trace sequential NOE walks between base protons and anomeric protons (H1'), and assigned all of them.