Inhibitory effects of ZSTK474, a phosphatidylinositol 3-kinase inhibitor, on adjuvant-induced arthritis in rats.

OBJECTIVE: We examined the effects of ZSTK474, a phosphatidylinositol 3-kinase (PI3K) inhibitor, on adjuvant-induced arthritis (AIA). METHODS: AIA was induced in Lewis rats by subcutaneous administration of Freund's complete adjuvant at the base of the tail on day 0. ZSTK474 was orally administered once daily from day 10. The severity of AIA was assessed by measuring the hind paw volume. The number of lymphocytes in inguinal lymph nodes (ILN) was determined by flow cytometry. The in vitro effects of ZSTK474 on the cell proliferation, and the cytokines and prostaglandin E(2) (PGE(2)) production were evaluated by BrdU method, ELISA and cytometric beads array. RESULTS: ZSTK474 ameliorated the progression of AIA. The temporary increases in the number of T cells in ILN, which occurred along with the appearance of arthritis, were inhibited in the ZSTK474-treated groups. In vitro studies revealed that ZSTK474 inhibited the production of IFNγ and IL-17 in concanavalin A-activated T cells. In vitro studies further revealed that ZSTK474 inhibited the proliferation and PGE(2) production by fibroblast-like synovial cells (FLS). CONCLUSION: ZSTK474 demonstrated prophylactic efficacy in a rat model of rheumatoid arthritis (RA) through inhibition of T cell and FLS functions. It was suggested that the inhibitors of PI3K have therapeutic potential for RA.

Human change and adaptation in Antarctica: Psychological research on Antarctic wintering-over at Syowa station.

An Antarctic wintering-over station is a unique environment, as a small isolated society facing extreme survival margins. Psychological surveys have been done over ten years, including the Baum test, the Positive and Negative Affect Schedule (PANAS), the Coping Orientation to Problems Experienced (COPE), Subjective Health Complaints Inventory (SHC), the Two-Sided Personality Scale (TSPS) and medical consultations in Syowa Station, a Japanese Antarctic station to reveal the mental status of team members. Team members experienced fewer physical health risks in Antarctica than in Japan. Wintering-over team members reinterpreted situations positively and accepted their environment, sought instrumental social support, planned ahead, and used active coping skills and humour to overcome difficulties. They did not act out emotionally or deny problems. Individuals exhibited two types of coping, either stability through maintaining a previous lifestyle or flexible adjustment to a new way of life. Positive affect remained constant during the wintering-over period. In living through a harsh reality, team members drew support from the subjective feelings of an "internal relationship" with home or family in their minds. Thus, an Antarctic wintering-over station is an ideal isolated environment for psychological surveys, which can help understand future space travel and group managements in everyday societies.

A Novel PEGylation Method for Improving the Pharmacokinetic Properties of Anti-Interleukin-17A RNA Aptamers.

The obstacles to the development of therapeutic aptamers for systemic inflammatory diseases, such as nuclease degradation and renal clearance, have not been fully overcome. Here, we report a novel PEGylation method, sbC-PEGylation, which improves the pharmacokinetic properties of RNA aptamers that act against interleukin-17A (IL-17A) in mice and monkeys. sbC-PEGylated aptamers were synthesized by coupling the symmetrical branching molecule 2-cyanoethyl-N,N-diisopropyl phosphoroamidite to the 5' end of the aptamer, before conjugating two polyethylene glycol (PEG) molecules to the aptamer. Pharmacokinetic studies showed that compared with conventionally PEGylated aptamers, the sbC-PEGylated aptamer exhibited excellent stability in the blood circulation of mice and monkeys. In addition, one of the sbC-PEGylated aptamers, 17M-382, inhibited the interleukin-6 (IL-6) production induced by IL-17A in NIH3T3 cells in a concentration-dependent manner, and the half-maximal inhibitory concentration of sbC-PEGylated 17M-382 was two times lower than that of non-PEGylated 17M-382. Furthermore, the intraperitoneal administration of sbC-PEGylated 17M-382 significantly inhibited the IL-6 production induced by IL-17A in a mouse air pouch model. Our findings suggest that the novel PEGylation method described in this study, sbC-PEGylation, could be used to develop anti-IL-17A aptamers as a therapeutic option for systemic inflammatory disease.

Characteristics of Staphylococcus intermedius isolates from diseased and healthy dogs.

Staphylococcus intermedius isolates from diseased and healthy dogs were examined for production of extracellular enzymes and toxins, and phage patterns. There were no significant differences between the two groups of isolates in the production rates of DNase, protease, lipase, gelatinase, hyaluronidase, hemolysins, protein A, and TSST-1, or in phage patterns. But the production rate of enterotoxins in isolates from diseased dogs was significantly higher than that in isolates from healthy dogs. PFGE analysis was performed with isolates from different body sites in individual dogs. In 3 of 6 healthy dogs, identical PFGE patterns were seen in isolates from the nares, external auditory meatus or skin. The remaining 3 dogs yielded isolates of different patterns. In 4 of 6 diseased dogs, identical patterns were seen in isolates from lesions as well as from the other normal sites.

Isolation and species distribution of staphylococci from animal and human skin.

From April 1999 to December 2000, a survey was made on the distribution of Staphylococcus species on the skin of 7 kinds of animals and humans. Staphylococci were isolated from 12 (100%) of 12 pigs, 17 (89.5%) of 19 horses, 30 (100%) of 30 cows, 73 (90.1%) of 81 chickens, 10 (40%) of 25 dogs, 23 (76.7%) of 30 laboratory mice, 20 (52.6%) of 38 pigeons, and 80 (88.9%) of 90 human beings. The predominant staphylococci isolated from a variety of animal species were novobiocin-resistant species, S. xylosus and S. sciuri regardless of the animal host species. The novobiocin-resistant species including S. xylosus and S. sciuri were only occasionally isolated from human skin. The predominant staphylococci found on human skin were novobiocin-sensitive species, S. epidermidis (63.8%), followed by S. warneri (28.8%) and S. hominis (13.8%). The results suggest that the staphylococcal flora inhabiting animal skin are different from those of human skin in regard to the predominant species isolated. In this study, we used pulsed-field gel electrophoresis to examine the chromosomal polymorphisms of S. epidermidis isolated most frequently from human skin. Strains of S. epidermidis showed the greatest genomic diversity in their fragment patterns.

Coronary artery revascularization before peripheral vascular surgery in patients with peripheral arterial disease.

The prevalence of coronary artery disease (CAD) in patients with peripheral arterial disease (PAD) approaches 50%. The incidence of perioperative cardiac complications is high in patients undergoing peripheral vascular surgery (PVS). However, the long-term efficacy of coronary artery revascularization in patients with PAD prior to PVS remains controversial. We retrospectively analyzed the long-term outcomes of 114 patients who underwent elective PVS. Coronary angiography (CAG) was performed routinely in all patients prior to the surgery. Preoperative CAG revealed CAD in 52 patients and no CAD in 62 patients (No-CAD group). Of the 52 patients with CAD, 15 patients with effort angina (more than CCS class 2), myocardial ischemia proven by pharmacological stress scintigraphy, and/or multivessel disease with impaired left ventricular systolic function underwent coronary revascularization (CAD-R group). The remaining 37 patients with CAD were treated medically (CAD-M group). The rates of postoperative events within 30 days were 26.7% in the CAD-R group, 10.8% in the CAD-M group, and 8.1% in the No-CAD group, respectively (P = 0.13), and the rates of long-term cardiovascular events were 33.3, 21.6, and 21.0%, respectively (P = 0.60). Therefore, the acute and long-term clinical outcomes in the CAD-R group were comparable with the other groups. In this study, the patients with CAD who showed inducible myocardial ischemia and/or multivessel disease with impaired left ventricular systolic function underwent coronary artery revascularization prior to the elective PVS. The patients who underwent coronary revascularization prior to the PVS exhibited comparable long-term outcomes compared with the patients who showed CAD but without inducible myocardial ischemia and with those without CAD. These data suggest that the evaluation of CAD and myocardial ischemia to determine the therapeutic strategy for CAD before elective PVS would be needed.

Coronary perforation during percutaneous coronary intervention.

Coronary perforation is an undesirable complication during percutaneous coronary intervention (PCI). We reviewed the cases of overt coronary perforation in our institute and analyzed their clinical backgrounds, the characteristics of the target lesion, management, and clinical outcomes. Between 1991 and 2005, we experienced 12 cases (0.35%) of coronary perforation in a total of 3415 PCI procedures. The perforation occurred during the use of debulking devices in 3 cases, immediately after stenting in 2, immediately after postdilatation of the stent in 2, and during wiring in 3 cases. Restoration was attempted by long inflation of a balloon in 7 cases, implantation of a covered stent graft in 1, and emergency surgical repair in 1 case. Subsequent cardiac tamponade occurred in 3 patients who required pericardiocentesis, and 1 patient died due to congestive heart failure. Administration of protamine was effective in stopping the bleeding in 6 patients, whereas continuation of antiplatelet therapy resulted in no overt rebleeding. Coronary perforation during PCI is a rare complication but is associated with significant morbidity and mortality. Intravenous administration of protamine is effective when it is used in conjunction with nonsurgical devices for initial management of perforation.

Cloning of a nitrate reductase inactivator (NRI) cDNA from Spinacia oleracea L. and expression of mRNA and protein of NRI in cultured spinach cells.

The spinach ( Spinacia oleracea L. (cv. Hoyo) nitrate reductase inactivator (NRI) is a novel protein that irreversibly inactivates NR. Using degenerate primers based on an N-terminal amino acid sequence of NRI purified from spinach leaves and a cDNA library, we isolated a full-length NRI cDNA from spinach that contains an open reading frame encoding 479 amino acid residues. This protein shares 67.4% and 51.1-68.3% amino acid sequence similarities with a nucleotide pyrophosphatase (EC 3.6.1.9) from rice and three types of the nucleotide pyrophosphatase-like protein from Arabidopsis thaliana, respectively. Immunoblot analysis revealed that NRI was constitutively expressed in suspension-cultured spinach cells; however, its expression level is quite low in 1-day-subcultured cells. Moreover, northern blot analysis indicated that this expression was regulated at the mRNA level. These results suggest that NRI functions in mature cells.