Candidatus Bartonella merieuxii,a Potential New Zoonotic Bartonella Species in Canids from Iraq

Bartonellae are emerging vector-borne pathogens infecting erythrocytes and endothelial cells of various domestic and wild mammals. Blood samples were collected from domestic and wild canids in Iraq under the United States Army zoonotic disease surveillance program. Serology was performed using an indirect immunofluorescent antibody test for B. henselae, B. clarridgeiae, B. vinsonii subsp. berkhoffii and B. bovis. Overall seroprevalence was 47.4% in dogs (n = 97), 40.4% in jackals (n = 57) and 12.8% in red foxes (n = 39). Bartonella species DNA was amplified from whole blood and representative strains were sequenced. DNA of a new Bartonella species similar to but distinct from B. bovis, was amplified from 37.1% of the dogs and 12.3% of the jackals. B. vinsonii subsp. berkhoffii was also amplified from one jackal and no Bartonella DNA was amplified from foxes. Adjusting for age, the odds of dogs being Bartonella PCR positive were 11.94 times higher than for wild canids (95% CI: 4.55–31.35), suggesting their role as reservoir for this new Bartonella species. This study reports on the prevalence of Bartonella species in domestic and wild canids of Iraq and provides the first detection of Bartonella in jackals. We propose Candidatus Bartonella merieuxii for this new Bartonella species. Most of the Bartonella species identified in sick dogs are also pathogenic for humans. Therefore, seroprevalence in Iraqi dog owners and bacteremia in Iraqi people with unexplained fever or culture negative endocarditis requires further investigation as well as in United States military personnel who were stationed in Iraq. Finally, it will also be essential to test any dog brought back from Iraq to the USA for presence of Bartonella bacteremia to prevent any accidental introduction of a new Bartonella species to the New World.     

Establishment of strongly neutralizing monoclonal antibody to human interleukin-6 and its epitope analysis

Three monoclonal antibodies against human interleukin-6 were established and characterized. One antibody was shown to strongly neutralize both the Ig-inducing and hybridoma/plasmacytoma growth activity of interleukin-6. The results of its epitope analysis using protease treated interleukin-6 and immobilized antibody indicated that this neutralizing antibody binds to a peptide corresponding to Leu151-Lys171 of interleukin-6 molecule. Further analysis using synthetic peptides showed that a shorter peptide corresponding to Ala153-Thr162 can also inhibit the binding of the antibody to interleukin-6. These results suggest that this carboxyl-terminal region plays a crucial role in interleukin-6 functions.     

Isolation of alpha-connectin, an elastic protein, from rabbit skeletal muscle

alpha-Connectin (also called titin 1) has been isolated from rabbit back muscle. Myofibrils were well washed with 5 mM NaHCO3 and then extracted with 0.2 M sodium phosphate, pH 7.0. The extract was dialyzed against 0.1 M potassium phosphate, pH 7.0, to sediment myosin. The supernatant, adjusted to 0.18 M potassium phosphate, pH 7.0, and 4 M urea, was subjected to DEAE Toyopearl column chromatography. beta-Connectin was eluted in the flow-through fraction and alpha-connectin was eluted at around 0.1 M NaCl, when a 0 to 0.25 M NaCl gradient was applied. The separated alpha-connectin was dialyzed against 0.2 M potassium phosphate, pH 7.0. The resultant alpha-connectin showed the same mobility as that in an SDS extract of rabbit back muscle on SDS gel electrophoresis using 1.8% polyacrylamide gels. A monoclonal antibody against chicken breast muscle beta-connectin reacted with the alpha-connectin isolated from rabbit back muscle.     

Differences in ventilatory responses to hypoxia and hypercapnia between normal and judo athletes with moderate obesity

Hypercapnic and hypoxic ventilatory sensitivities were compared in twenty-one judoists and 24 control subjects with similar degrees of moderate obesity. Data from ten non-obese control subjects were also included as a reference. Mean body weight (BW) and % of ideal body weight in the judoists and the obese and non-obese controls were 100 +/- 14.8, 94.4 +/- 5.3 and 63.4 +/- 6.1 (mean +/- SD) kg, and 142.3 +/- 16.7, 142.2 +/- 12.9 and 98.4 +/- 10.7%, respectively. Mean body fat in the judoists was 16.2 +/- 13.9%, being 25.3 +/- 7.7% in the obese control group, the difference being significant (p less than 0.01). Hypercapnic sensitivities in terms of the CO2 ventilatory response slope (S) and its normalized value for 70 kg BW (SN) of the obese controls were higher than the judoists. These findings were also verified by the CO2-occlusion pressure responses. S and SN in the obese controls were significantly correlated with BW and % body fat. However, no positive correlation was found between BW and S or SN in the judoists as well as between lean body mass and S or SN in the obese control. Hypoxic sensitivity in terms of the PETO2-ventilation hyperbola slope (A) and its normalized value (AN) in the obese control was significantly higher than the non-obese control, but the difference from the judoists was not significant. A and AN were found to increase with increasing % body fat in both judoists and obese controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation of Bartonella henselae and Two New Bartonella Subspecies,Bartonella koehlerae Subspecies boulouisii subsp. nov. and Bartonella koehlerae Subspecies bothieri subsp. nov. from Free-Ranging Californian Mountain Lions and Bobcats

Domestic cats are the natural reservoir of Bartonella henselae, B. clarridgeiae and B. koehlerae. To determine the role of wild felids in the epidemiology of Bartonella infections, blood was collected from 14 free-ranging California mountain lions (Puma concolor) and 19 bobcats (Lynx rufus). Bartonella spp. were isolated from four (29%) mountain lions and seven (37%) bobcats. These isolates were characterized using growth characteristics, biochemical reactions, molecular techniques, including PCR-RFLP of selected genes or interspacer region, pulsed-field gel electrophoresis (PFGE), partial sequencing of several genes, and DNA-DNA hybridization. Two isolates were identical to B. henselae genotype II. All other isolates were distinguished from B. henselae and B. koehlerae by PCR-RFLP of the gltA gene using endonucleases HhaI, TaqI and AciI, with the latter two discriminating between the mountain lion and the bobcat isolates. These two novel isolates displayed specific PFGE profiles distinct from B. henselae, B. koehlerae and B. clarridgeiae. Sequences of amplified gene fragments from the three mountain lion and six bobcat isolates were closely related to, but distinct from, B. henselae and B. koehlerae. Finally, DNA-DNA hybridization studies demonstrated that the mountain lion and bobcat strains are most closely related to B. koehlerae. We propose naming the mountain lion isolates B. koehlerae subsp. boulouisii subsp. nov. (type strain: L-42-94), and the bobcat isolates B. koehlerae subsp. bothieri subsp. nov. (type strain: L-17-96), and to emend B. koehlerae as B. koehlerae subsp. koehlerae. The mode of transmission and the zoonotic potential of these new Bartonella subspecies remain to be determined.     

Influence of Molecular Structure on O2-Binding Properties and Blood Circulation of Hemoglobin?Albumin Clusters

A hemoglobin wrapped covalently by three human serum albumins, a Hb-HSA₃ cluster, is an artificial O₂-carrier with the potential to function as a red blood cell substitute. This paper describes the synthesis and O₂-binding properties of new hemoglobin‒albumin clusters (i) bearing four HSA units at the periphery (Hb-HSA₄, large-size variant) and (ii) containing an intramolecularly crosslinked Hb in the center (XLHb-HSA₃, high O₂-affinity variant). Dynamic light scattering measurements revealed that the Hb-HSA₄ diameter is greater than that of either Hb-HSA₃ or XLHb-HSA₃. The XLHb-HSA₃ showed moderately high O₂-affinity compared to the others because of the chemical linkage between the Cys-93(β) residues in Hb. Furthermore, the blood circulation behavior of ¹²⁵I-labeled clusters was investigated by assay of blood retention and tissue distribution after intravenous administration into anesthetized rats. The XLHb-HSA₃ was metabolized faster than Hb-HSA₃ and Hb-HSA₄. Results suggest that the molecular structure of the protein cluster is a factor that can influence in vivo circulation behavior.     

Temperature response and photoinhibition investigated by chlorophyll fluorescence measurements for four distinct species of dipterocarp trees

The effects of strong light in combination with elevated temperatures on the photosynthetic system were examined in 4 dipterocarp tree species with ecologically different habitats. The 4 dipterocarp tree species were: Shorea platyclados originated from upper dipterocarp forests, Shorea parvifolia– lowland and hill dipterocarp forests, Shorea assamica– lowland dipterocarp forests, and Dipterocarpus oblongifolius– riparian fringes. S. platyclados and D. oblongifolius have higher growth and survival rates in open sites than S. parvifolia and S. assamica. Tolerance of high temperature among the species was assessed by determining the critical temperatures (T_c) at which the minimal fluorescence (F_o) began to rise sharply. This was measured by exposing plants to an increasing temperature of about 1°C min^?1. The intrinsic thermotolerance of the thylakoid membrane appears to be the highest for D. oblongifolius (T_c=46.4°C), intermediate for S. platyclados (45.7°C), and lowest for S. parvifolia and S. assamica (45.2 and 45.3°C, respectively). The temperature‐dependent efficiency of PSII electron transport (ΔF/F′_m), photochemical quenching (q_P), and the efficiency of light capture of open PSII (F′_v/F′_m) were measured at the photosynthetic steady state at least 10 min after the light exposure (180 μmol m^?2 s^?1 PFD). Stable temperature responses of ΔF/F′_m and q_P were observed in S. platyclados and D. oblongifolius, while those in S. parvifolia and S. assamica were more temperature‐dependent and severely affected at 45°C. Little difference was observed in temperature‐dependent F′_v/F′_m among species. Photoinhibitory light exposure (1600 μmol m^?2 s^?1 PFD) for 2 h at 40°C had little effect on the recovery kinetics from photoinhibition of S. platyclados and D. oblongifolius compared with those at 35°C. In contrast, the recovery from photoinhibition was retarded in S. parvifolia and S. assamica. These findings suggest that even at 40°C, a temperature below T_c, an exposure to strong light exacerbated photoinhibition in S. parvifolia and S. assamica corresponding to the closure of PSII reaction centers, as indicated by the decrease in q_P at this temperature. Thus, S. platyclados and D. oblongifolius, which occur at uplands and riparian fringes with frequent disturbances, are suggested to have higher photosynthetic tolerance to elevated temperatures contributing to a circumvention of photoinhibition.     

DNA polymerases during tumor growth

DNA polymerase activity was studied as a function of stage of tumor growth and correlated with DNA synthesis measured by 3H-TdR uptake. Considerable variations in DNA synthesis activity occur at different growth stages and following host death. DNA alpha-polymerase activity did vary with growth stage in the ascites tumor. However, it did not have a clear correlation with DNA synthesis or with tumor growth. No striking fall in DNA polymerase enzyme levels occurred as the ascites tumor reached stationary phase in contrast to reports in some cell culture systems. A decrease occurred with advanced tumor stage and after host death. DNA beta-polymerase activity did not change with tumor growth stage.