The efficacy of intravenous injection of FK463, a novel water-soluble lipopeptide, was evaluated in mouse models of disseminated candidiasis and aspergillosis and was compared with those of fluconazole (FLCZ) and amphotericin B (AMPH-B). In the candidiasis model, FK463 significantly prolonged the survival of intravenously infected mice at doses of 0.125 mg/kg of body weight or higher. In disseminated candidiasis caused by Candida species, including FLCZ-resistant Candida albicans, FK463 exhibited an efficacy 1.4 to 18 times inferior to that of AMPH-B, with 50% effective doses (ED 50 s) ranging from 0.21 to 1.00 mg/kg and 0.06 to 0.26 mg/kg, respectively, and was much more active than FLCZ. The protective effect of FK463 was not obviously influenced by the fungal inoculum size, the starting time of the treatment, or the immunosuppressed status of the host. The reduction in efficacy was less than that observed with FLCZ or AMPH-B. The efficacy of FK463 was also evaluated in the disseminated candidiasis target organ assay and was compared with those of FLCZ and AMPH-B. Efficacies were evaluated on the basis of a comparison between the mean log 10 CFU in kidneys in the groups treated with antifungal agents and that in control group. A single dose of FK463 at 0.5 mg/kg or higher significantly reduced the viable counts in kidneys compared with the numbers of yeast cells before treatment, and its efficacy was comparable to that of AMPH-B, while FLCZ at 4 mg/kg showed only a suppressive effect on the growth of C. albicans in the kidneys. In the disseminated aspergillosis model, FK463 given at doses of 0.5 mg/kg or higher significantly prolonged the survival of mice infected intravenously with Aspergillus fumigatus conidia. The efficacy of FK463 was about 2 times inferior to that of AMPH-B, with ED 50 s ranging from 0.25 to 0.50 mg/kg and 0.11 to 0.29 mg/kg, respectively. These results indicate that FK463 may be a potent parenterally administered therapeutic agent for disseminated candidiasis and aspergillosis.People who have impaired immune systems are susceptible to fungal infections which can be life-threatening. Immune deficiencies resulting from AIDS, aggressive cancer treatment, the growing use of organ transplants, and other nosocomial situations have greatly increased the incidence of serious fungal infections (2,3,4,6) and have created a critical need for new, safe fungicidal agents that can be used to treat disseminated infections. Systemic mycoses are not easily diagnosed, and the patient usually has been infected for quite some time before symptoms appear. Thus, empiric therapy needs to begin immediately, but currently available treatments have problems with toxicity or resistance. Amphotericin B (AMPH-B) is the first-line therapy for systemic infections because of its broadspectrum and fungicidal activity. However, significant side effects limit its clinical utility to controlled intravenous administration (16). Lipid AMPH-B formulations have recently attracted much attention due to significantly lower toxic...

The profiles of the resonance line of potassium (KI 7698.98) in the flux spectrum of the Sun and Procyon were studied in detail using the multi-parameter fitting method, in order to extract information regarding the fundamental atomic parameters responsible for the formation of this line as well as to diagnose the nature of the velocity fields in their atmospheres. It was concluded from an analysis of the solar KI 7699 profile that neutral-hydrogen collisions are practically negligible compared to electron collisions in the rate equation of statistical equilibrium, and that an empirical correction to the Unsöld's standard (van der Waals effect) damping constant is Δ log C6 ≃ + 1.0 with the most consistent microturbulent velocity of ξ ≃ 0.8 km s−1 . Based upon these results, the atmospheric-velocity parameters of Procyon were determined to be ξ ≃ 1.4 km s−1, ζRT≃ 6.7 km s−1 (the radial-tangential macroturbulence), and ve sin i ≃ 3.3 km s−1 (the projected rotational velocity). The potassium abundances of the Sun and Procyon derived from this resonance line, which are significantly affected by the non-LTE effect amounting to ∼ 0.4 dex (Sun) and ∼ 0.7 dex (Procyon), turned out to be nearly the same and log εK ≃ 5.1. It also appears that the microturbulence tends to decrease with an increase in the atmospheric height of the Sun as well as of Procyon.

The cells of Blepharisma which possess red pigment (blepharismin) show step‐up photophobic response (temporal ciliary reversal induced by a sudden increase in light intensity). Bleaching of the cells by cold shock raised a threshold light intensity for the response, Oxidation of red pigment that produced blue pigment did not raise the threshold for the response. The action spectrum for the step‐up photophobic response of the cells which possess normal red pigment had peaks at about 580, 540 and 490 nm, a value which coincided with peaks of an absorption spectrum of the red pigment. The absorption spectrum of oxidized pigment (blue pigment) shifted 20 nm toward infrared light. The action spectrum for the response of the cells which possess blue pigment also shifted 20 nm toward infrared light. Results suggest that red pigment might be involved in the step‐up photophobic response. Key words. Blepharismin, ciliary reversal, photoreceptors, photoresponse.

We constructed the expression vector pSK-SCP containing the streptococcal exotoxin B gene (spe b) which expressed protease activity. We showed that the recombinant streptococcal pyogenic exotoxin B/streptococcal cysteine protease (rSPE B/SCP) was secreted into the culture supernatant of the transformant and retained its SCP activity, which was equivalent to or greater than that of the naturally occurring molecule. The secreted rSPE B/SCP induced histamine release and degranulation of the human mast cell line HMC-1. This study may contribute to the understanding of the pathogenic role of SPE B/SCP in streptococcal infection and streptococcal toxic shock syndrome.Streptococcal pyrogenic exotoxin B (SPE B), known as streptococcal cysteine protease (SPE B/SCP) is highly conserved among group A Streptococcus (GAS). The structural gene, spe b, is found among all GAS clinical isolates (20) and expressed in almost all strains (12). This protein is initially secreted as a 42-kDa precursor, zymogen, and is autocatalytically cleaved to a 28-kDa mature SPE B/SCP (7, 9). SPE B/SCP cleaves or degrades fibronectin and vitronectin (12), which function as human matrix proteins. It also cleaves plasma kininogen and releases kinin (10) and cleaves the interleukin-1␤ (IL-1␤) precursor to the active form of IL-1␤ (12). Furthermore, SPE B/SCP activates the human matrix metalloprotease that increases type IV collagen degradation (2). Finally, genetically inactivated SPE B/SCP is attenuated in the mouse lethality model (15, 16). These findings suggest that SPE B/SCP activates inflammatory responses in the host and plays a very important role in pathogenesis of infections.The SPE B/SCP gene was previously cloned, sequenced (1, 9, 18), and expressed in Escherichia coli (6,8,17,19). However, the secretion of the active form of the protease from E. coli and its activity relative to the native molecule have not been examined. In order to clarify this question, we constructed an expression vector including spe b and its predicted promoter regions for transformation into E. coli. As a result, recombinant SPE B/SCP (rSPE B/SCP), retaining cysteine protease activity, was obtained from the culture supernatant.GAS strain NZ131 (kindly provided by D. R. Martin, NewZealand Communicable Disease Center, Porirua) chromosomal DNA was used as the template. Two oligonucleotides (SPEBF0008, 5ЈGTGTCAACTAACCGTGTTATTG-3Ј; SPEBR1485, 5Ј-TGATCTGTGTCTGASTGGATACTT-3Ј) were designed based on the spe b sequence as reported by Hauser et al. (9) and used as primers. PCR was performed with 25 cycles (94°C for 30 s, 54°C for 30 s, and 75°C for 1.5 min) and pyrobest DNA polymerase (TaKaRa Biomedicals, Kyoto, Japan). In order to construct the expression vector pSK-SCP, a 1,469-bp fragment including spe b and its predicted promoter region was amplified and cloned via the SmaI site in pBluescript II SK(ϩ) plasmid vector. pSK-SCP was transformed into E. coli strain JM109. The rSPE B/SCP was induced with 2 mM isopropyl-␤-D-thiogalactopyranoside (IPTG; Wako Pure Chemical Co...

We previously purified Streptococcus mitis-derived human platelet aggregation factor (Sm-hPAF) from the culture supernatant of S. mitis strain Nm-65, isolated from the tooth surface of a patient with Kawasaki disease. Here we produced recombinant Sm-hPAF protein (rSm-hPAF) in Escherichia coli, to determine whether rSm-hPAF conserves its platelet aggregation activity. rSm-hPAF precursor (665 amino acids) shows up to 36-56% identity with the family of cholesterol-dependent cytolysins (CDCs), and rSm-hPAF displayed potent hemolytic activity toward mammalian erythrocytes, including human erythrocytes with platelet aggregation activity. The 162-amino acid amino-terminal domain of rSm-hPAF was found in no other CDCs except lectinolysin; this domain is homologous to a portion of pneumococcal fucolectin-related protein. Interestingly, suilysin (SLY) and pneumolysin (PLY) of CDCs also exhibit substantial human platelet aggregation activity, similar to rSm-hPAF, and the platelet aggregation by rSm-hPAF, SLY, and PLY was morphologically confirmed using light and electron microscopy.