Clearance of Theileria sergenti-infected bovine red blood cells in severe combined immune deficiency mice

Clearance of Theileria sergenti-infected bovine red blood cells (Bo-RBCs) from the blood circulation of severe combined immune deficiency (SCID) mice was studied to help understand the mechanisms of anemia developing in cattle infected with T. sergenti. For the clearance test, Bo-RBC samples having 2%, 58%, and 76% parasitemia and, as a control, parasite-free Bo-RBCs were prepared in the Bo-RBC-SCID mouse model. The T. sergenti-infected Bo-RBCs and the uninfected control Bo-RBCs were separately labeled with two, green and red, fluorescent dyes, mixed together, and injected intravenously into SCID mice. The blood samples collected at various time points were observed under a fluorescent microscope, and the numbers of green and red fluorescing RBCs were counted differentially to determine the clearance rates of T. sergenti-infected and uninfected Bo-RBCs. This test clearly demonstrated that the Bo-RBC samples having higher parasitemias were cleared faster from the blood circulation of SCID mice. The results suggest that the intravascular clearance system in SCID mice may have a mechanism by which T. sergenti-parasitized and non-parasitized Bo-RBCs are recognized and cleared differentially.     

Temperature and metal ions-dependent activity of the family I inorganic pyrophosphatase from the swine roundworm Ascaris suum

Temperature dependence, heat stability and metal ions-dependent activity were examined on the Family I inorganic pyrophosphatase (PPase) recently identified from Ascaris suum. Recombinant A. suum PPase (rAsPPase) showed an optimal activity at 55 degrees C. The rAsPPase was heat stable at 40 degrees C in the absence of added Mg(2+) and at 50 degrees C in its presence. The enzyme required divalent metal ions for its activity. The preferences for the metal ions (5 mM concentration) were in the order: Mg(2+)> Co(2+)> Cu(2+)> Fe(2+)> Zn(2+)> Mn(2+). On the contrary, enzyme activity was inhibited by Ca(2+). These findings suggest that catalytic features of AsPPase are consistent with the Family I PPases reported from a wide range of organisms.     

Evaluation of QT interval prolongation in dogs with heart failure

Comparison of the QT interval and corrected QT interval values that were calculated by the methods of Bazett (QTc1) and Fridericia (QTc2) were made between dogs with or without cardiac diseases to determine the influence of the QT interval on canine heart failure. Upon comparison of the measured values on ECG between the cardiac disease and non-cardiac disease groups, it was observed that the heart rate(HR) was significantly higher in the cardiac disease group than in the non-cardiac disease group, although the QT interval was similar in the two groups. The QTc1 and QTc2 were significantly longer in the cardiac disease group than in the non-cardiac disease group. With the progression of the New York Heart Association Class, the HR tended to increase. The QTc1 and QTc2 became significantly prolonged with the progression of heart failure. Nevertheless, because Bazett's correction formula is known to overcorrect when the HR is high, it was considered that the QTc1 was actually overcorrected by high HR with the progression of heart failure. The QTc2, on the other hand, was only slightly influenced by HR, suggesting that the prolongation was due to the progression of heart failure. These results suggest that the prolongation of QTc2 in cardiac disease reflects the substantial prolongation of the QT interval without the influence of HR. It is suggested that the QTc2 could be a useful parameter for assessing the degree of heart failure in dogs with cardiac disease.     

Genetic selection for resistance to mycoplasmal pneumonia of swine (MPS) in the Landrace line influences the expression of soluble factors in blood after MPS vaccine sensitization

We recently developed a Landrace line that is resistant to mycoplasmal pneumonia of swine (MPS) infection by genetic selection for five generations, and we reported that the immunophenotype of this line is different from that of the non‐selected line in terms of changes in peripheral blood leukocyte population after MPS vaccination. This study followed up previous findings demonstrating changes in soluble factors in blood, namely, hormones, Mycoplasma hyopneumoniae‐specific immunoglobulin G (IgG), and cytokines. These two lines were injected with MPS vaccine on days ?7 and 0 after blood sampling on those days, and blood samples were collected on days ?14, ?7, 0, 2, 7 and 14. We found changes in the levels of many hormones and cytokines in both lines. However, we found that only growth hormone (GH) and interferon (IFN)‐γ levels were statistically different between these two lines. GH concentration was reduced (day 0) and IFN‐γ concentration was increased (day 14) in the MPS‐selected line compared with the non‐selected line, despite unchanged IFN‐γ messenger RNA expression in blood cells. Although detailed mechanisms underlying these phenotypes remain unsolved, these traits would be useful to improve MPS resistance in pig production and provide an insight into MPS infection.     

Murine hepatitis virus: effect on liver RNA

After infection of mice with hepatitis virus MHV3, the RNA in the liver undergoes changes. The fraction extracted with phenol at 0 degrees C does not alter. However, the fraction extracted with hot phenol at elevated pH (60 degrees C, pH 8.3) shows a 16S peak on sucrose-density-gradient centrifugation. This fraction shows actinomycin D-resistant incorporation of C(14)-orotic acid in infected but not in control livers-possible evidence of the RNA nature of MHV3.     

Structures of glycoprotein Ibalpha and its complex with von Willebrand factor A1 domain

Transient interactions of platelet-receptor glycoprotein Ibalpha (GpIbalpha) and the plasma protein von Willebrand factor (VWF) reduce platelet velocity at sites of vascular damage and play a role in haemostasis and thrombosis. Here we present structures of the GpIbalpha amino-terminal domain and its complex with the VWF domain A1. In the complex, GpIbalpha wraps around one side of A1, providing two contact areas bridged by an area of solvated charge interaction. The structures explain the effects of gain-of-function mutations related to bleeding disorders and provide a model for shear-induced activation. These detailed insights into the initial interactions in platelet adhesion are relevant to the development of antithrombotic drugs.     

Effects of tea polyphenols on the invasion and matrix metalloproteinases activities of human fibrosarcoma HT1080 cells

The effects of tea polyphenols on the invasion of highly metastatic human fibrosarcoma HT1080 cells through a monolayer of human umbilical vein endothelial cells (HUVECs) and the accompanying basal membrane were investigated. Among the tea polyphenols tested, epicatechin gallate (ECg), epigallocatechin gallate (EGCg), and theaflavin strongly suppressed the invasion of HT1080 cells into the monolayer of HUVECs/gelatin membrane, whereas epicatechin, epigallocatechin, tea flavonols, tea flavones, and gallate derivatives had no effect. Both theaflavin-digallate and theasinensin D showed a weak invasion inhibitory effect. ECg significantly inhibited the invasion without cytotoxicity against cancer cells and HUVECs. Ester-type catechins (ECg and EGCg) and theaflavin strongly suppressed the gelatin degradation mediated by matrix metalloproteinase (MMP) 2 and MMP-9, which were secreted into the conditioned medium of HT1080 cells. In conclusion, among the tea polyphenols tested, ECg was considered to be the agent with the most potential antimetastasis activity because it inhibited invasion in the absence of cytotoxicity.     

Simple sequence repeat markers linked to a major QTL controlling pod shattering in soybean

Shattering of soybean pods prior to harvest leads to a reduction in yield. In order to identify simple sequence repeat (SSR) markers linked to quantitative trait loci (QTLs) conditioning pod shattering, QTL analysis was conducted using an recombinant inbred line (RIL) population segregating for this trait. The degrees of pod‐shattering resistance were evaluated by heat treatment applied to pods harvested from plants in the field and in a growth chamber. Composite interval mapping identified one major QTL between SSR markers Sat_093 and Sat_366 on linkage group J for both environments. The position and the effect of this QTL were confirmed in an F₂ population derived from a cross between the pod shattering‐susceptible parental cultivar and a pod shattering‐resistant RIL. The SSR markers linked to the major QTL will be useful for marker‐assisted selection in soybean‐breeding programmes.     

Characterization of depth-related changes and site-specific differences of microbial communities in marine sediments using quinone profiles

ABSTRACT:   Respiratory quinone compositions were analyzed by a high-performance liquid chromatography to characterize the depth-related changes and site-specific differences of microbial communities in marine sediments. Two deep-sea sediment samples and one coastal sediment sample were investigated from three sites on the coast of Japan, Sagami Bay, Suruga Bay and Tokyo Bay. Although depth-related changes in microbial community structures were observed, site-specific differences appeared to have greater influence on overall community structures. A variety of quinone homologs was commonly identified at all sampling sites and depths examined, but a few minor quinone fractions, mainly derived from Actinobacteria , were detected only at specific sampling sites. Methylmenaquinone-7 (MMK-7) was the major component throughout the examined depths in Sagami Bay and Tokyo Bay. Most abundant quinone homologs changed with each depth in Suruga Bay. Menaquinone-6 (MK-6) predominated in the 0 to 2-cm layer (19%). Below 0–2 cm, the most abundant homolog in each sampling depth was phylloquinone (K₁; 13%, 4–6 cm), MK-8 (19%, 8–10 cm) and MK-7 (13%, 16–18 cm). The microbial respiratory quinone profiling method shown here, successfully demonstrates the usefulness of this approach to characterize microbial communities in marine sediments.