Evaluation of Francisella tularensis ΔpdpC as a candidate live attenuated vaccine against respiratory challenge by a virulent SCHU P9 strain of Francisella tularensis in a C57BL/6J mouse model

Francisella tularensis, which causes tularemia, is an intracellular gram‐negative bacterium. F. tularensis has received significant attention in recent decades because of its history as a biological weapon. Thus, development of novel vaccines against tularemia has been an important goal. The attenuated F. tularensis strain ΔpdpC, in which the pathogenicity determinant protein C gene (pdpC) has been disrupted by TargeTron mutagenesis, was investigated as a potential vaccine candidate for tularemia in the present study. C57BL/6J mice immunized s.c. with 1 × 10⁶ CFUs of ΔpdpC were challenged intranasally with 100× the median lethal dose (LD₅₀) of a virulent SCHU P9 strain 21 days post immunization. Protection against this challenge was achieved in 38% of immunized C57BL/6J mice administered 100 LD₅₀ of this strain. Conversely, all unimmunized mice succumbed to death 6 days post challenge. Survival rates were significantly higher in vaccinated than in unimmunized mice. In addition, ΔpdpC was passaged serially in mice to confirm its stable attenuation. Low bacterial loads persisted in mouse spleens during the first to tenth passages. No statistically significant changes in the number of CFUs were observed during in vivo passage of ΔpdpC. The inserted intron sequences for disrupting pdpC were completely maintained even after the tenth passage in mice. Considering the stable attenuation and intron sequences, it is suggested that ΔpdpC is a promising tularemia vaccine candidate.     

Biosorption of Cr, Cu and Al bySargassum biomass

The biosorption and desorption of Cr, Cu and Al were carried out using brown marine algaeSargassum fluitans biomass, known as the good biosorbent of heavy metals. The content of alginate bound to light metals could be changed by physical and chemical pretreatment. The maximum uptake of Cr, Cu and Al was independent of the alginate content. The maximum uptake of Al was two times(mole basis) than those of Cu and Cr. The aluminum-alginate complex was found in the sorption solution of raw and protonated biomass. Most of Cu, Al and light metals sorbed in the biomass were eluted at pH 1.1. However, only 5 to 10% of Cr sorbed was eluted at pH 1.1. The stoichometric ion exchange between Cu and Ca ion was observed on Cu biosorption with Ca-loaded biomass. A part of Cr ion was bound to biomass as Cr(OH)₂ ⁺ or Cr(OH)²⁺. Al was also bound to biomass as multi-valence ion and interfered with the desorbed Ca ion. The behavior of rawS. fluitans in ten consecutive sorption-desorption cycles has been investigated in a packed bed flow-through-column during a continuous removal of copper from a 35 mg/L aqueous solution at pH 5. The eluant used was a 1%(w/v) CaCl/HCl solution at pH3.     

Biomass accumulations and the distribution of nitrogen and phosphorus within three<Emphasis Type="Italic">Quercus acutissima</Emphasis> stands in central Korea

Above- and belowground biomass and nitrogen (N) and phosphorus (P) distribution within threeQuercus acutissima stands were investigated in central Korea. The average age (year) and diameter at breast height (DBH, cm) were 10.8 and 7.9 for Stand 1, 38.2 and 17.1 for Stand 2, and 44.0 and 20.7 for Stand 3, respectively. Fifteen trees were destructively harvested for dimension analysis of component biomass (stem wood, stem bark, foliage, branches, and roots) plus N and P concentrations. Total biomass (t ha^-1) was 88.7 for Stand 1, 154.9 for Stand 2, and 278.1 for Stand 3 while N and P contents in all tree components (kg ha^-1) were 483.3 and 52.2, 697.1 and 55.0, and 1113.9 and 83.7. Nitrogen concentrations were highest in the foliage, followed by the stem bark, branches or roots, and stem wood. In contrast, P concentrations were greatest in the roots, then foliage, branches, stem bark, and stem wood. In general, N and P concentrations in these components significantly decreased with tree age and DBH, while N and P contents significantly increased with age and size. These relationships were stronger for size than for age. Our current data could be utilized to estimate N and P budgets for silvicultural practices, including fertilization, thinning, and harvesting.     

Study on the ophthalmic diseases in ICR mice and BALB/c mice.

In pharmaceutical companies and research institutes, many toxicity tests are performed with laboratory animals. This study was performed to produce reference data for eye toxicity tests and to investigate the ophthalmic diseases of 408 ICR mice and 119 BALB/c mice, which are commonly used as subjects in toxicity tests. The experimental animals without clinical disorders were selected regardless of sex. The ophthalmic diseases were examined by using special ophthalmic instruments: direct ophthalmoscope, indirect ophthalmoscope, slit-lamp biomicroscope and focal illuminator. The most prevalent ocular variation within normal limits was hyaloid vessel remnant (ICR mice, 28.2%; BALB/c mice, 31.9%) and the incidence gradually decreased with age. The ocular diseases found in ICR mice were retinal degeneration (9.8%), corneal scar (4.2%), focal cataract (2.2%), anisocoria (1.2%), corneal ulcer (0.2%) and uveitis (0.2%). In BALB/c mice, corneal scar (9.2%), focal cataract (1.7%) and corneal ulcer (0.8%) were the ocular diseases found.     

The dynamic regulation of myocardial oxidative phosphorylation: Analysis of the response time of oxygen consumption

Although usually steady-state fluxes and metabolite levels are assessed for the study of metabolic regulation, much can be learned from studying the transient response during quick changes of an input to the system. To this end we study the transient response of O₂ consumption in the heart during steps in heart rate. The time course is characterized by the mean response time of O₂ consumption which is the first statistical moment of the impulse response function of the system (for mono-exponential responses equal to the time constant). The time course of O₂ uptake during quick changes is measured with O₂ electrodes in the arterial perfusate and venous effluent of the heart, but the venous signal is delayed with respect to O₂ consumption in the mitochondria due to O₂ diffusion and vascular transport. We correct for this transport delay by using the mass balance of O₂, with all terms (e.g. O₂ consumption and vascular O₂ transport) taken as function of time. Integration of this mass balance over the duration of the response yields a relation between the mean transit time for O₂ and changes in cardiac O₂ content. Experimental data on the response times of venous [O₂] during step changes in arterial [O₂] or in perfusion flow are used to calculate the transport time between mitochondria and the venous O₂ electrode. By subtracting the transport time from the response time measured in the venous outflow the mean response time of mitochondrial O₂ consumption (t_mito) to the step in heart rate is obtained.In isolated rabbit heart we found that t_mito to heart rate steps is 4-12 s at 37°C. This means that oxidative phosphorylation responds to changing ATP hydrolysis with some delay, so that the phosphocreatine levels in the heart must be decreased, at least in the early stages after an increase in cardiac ATP hydrolysis. Changes in ADP and inorganic phosphate (P_i) thus play a role in regulating the dynamic adaptation of oxidative phosphorylation, although most steady state NMR measurements in the heart had suggested that ADP and P_i do not change. Indeed, we found with ³¹P-NMR spectroscopy that phosphocreatine (PCr) and P_i change in the first seconds after a quick change in ATP hydrolysis, but remarkably they do this significantly faster (time constant ~2.5 s) than mitochondrial O₂ consumption (time constant 12 s). Although it is quite likely that other factors besides ADP and P_i regulate cardiac oxidative phosphorylation, a fascinating alternative explanation is that the first changes in PCr measured with NMR spectroscopy took exclusively place in or near the myofibrils, and that a metabolic wave must then travel with some delay to the mitochondria to stimulate oxidative phosphorylation. The t_mito slows with falling temperature, intracellular acidosis, and sometimes also during reperfusion following ischemia and with decreased mitochondrial aerobic capacity. In conclusion, the study of the dynamic adaptation of cardiac oxidative phosphorylation to demand using the mean response time of cardiac mitochondrial O₂ consumption is a very valuable tool to investigate the regulation of cardiac mitochondrial energy metabolism in health and disease.     

Follow-Up Study of Hypervariable Region Sequences of the Hepatitis C Virus (HCV) Genome in an Infant with Delayed Anti-HCV Antibody Responses

An infant born prematurely and infected with hepatitis C virus (HCV) one month after birth was followed for 4.5 years. The patient did not produce detectable anti-HCV antibodies until two years after the onset of hepatitis. Before seroconversion, a single clone of HCV, as determined by quasispecies of the hypervariable region (HVR) of the HCV genome, was almost exclusively found in the serum. After seroconversion, however, another distinct lineage of HCV clones replaced it within half a year. As HCV infection persisted further in the presence of anti-HCV antibodies, many derivatives of both sequence lineages emerged to exhibit the typical quasispecies feature of HVR sequences. Neither seroconversion nor the changes in HVR sequences influenced the serum aminotransferase titers.     

Serological Typing of 31 Achromogenic and 40 Melanogenic Pseudomonas aeruginosa Strains

Thirty-one achromogenic and 40 melanogenic Pseudomonas aeruginosa strains were studied with 10 monovalent typing sera (3). Twenty-one of the achromogenic (67.7%) and seven of the melanogenic (17.5%) strains were agglutinated by one of the 10 typing sera. Ten achromogenic and 33 melanogenic strains were not agglutinated by any of the 10 typing sera. As far as this set of antisera is concerned, the typability of achromogenic and melanogenic P. aeruginosa strains appears to be much lower than that of the chromogenic, nonmelanogenic strains of the species reported previously.