Determination of the vector species of tsutsugamushi disease in Korea

In order to determine the vector species of tsutsugamushi disease in Korea, chiggers were individually dissected, and internal contents were tested for Rickettsia tsutsugamushi organisms by means of indirect FA test, and each exoskeleton was mounted on slide for identification. Among 4,142 chiggers collected from 48 Apodemus agrarius at nine different localities during the period of July-November, 1989, 990 chiggers of 10 species of Trombiculidae were dissected and tested. Rickettsiae were confirmed in two Leptotrombidium pallidum larvae out of 447 tested, giving 0.4% of the infection rate. The chiggers of the other species tested were found negative.     

Molecular nature of Spemann's organizer: the role of the Xenopus homeobox gene goosecoid.

This study analyzes the function of the homeobox gene goosecoid in Xenopus development. First, we find that goosecoid mRNA distribution closely mimics the expected localization of organizer tissue in normal embryos as well as in those treated with LiCl and UV light. Second, goosecoid mRNA accumulation is induced by activin, even in the absence of protein synthesis. It is not affected by bFGF and is repressed by retinoic acid. Lastly, microinjection of goosecoid mRNA into the ventral side of Xenopus embryos, where goosecoid is normally absent, leads to the formation of an additional complete body axis, including head structures and abundant notochordal tissue. The results suggest that the goosecoid homeodomain protein plays a central role in executing Spemann's organizer phenomenon.     

Overexpression of a homeodomain protein confers axis-forming activity to uncommitted Xenopus embryonic cells.

The anteroposterior character of mesoderm induced by a peptide growth factor (XTC-MIF) was tested by transplantation into host Xenopus gastrulae. Both retinoic acid and a homeodomain protein were able to override the anteriorizing effect of the growth factor. Microinjection of a posteriorly expressed homeobox mRNA can respecify anteroposterior identity, transforming head mesoderm into tail-inducing mesoderm. Unexpectedly, overexpression of XIHbox 6 protein in the transplanted cells, without addition of growth factors, caused the formation of tail-like structures. The cells overexpressing XIHbox 6 were able to recruit cells from the host into the secondary axis. The results suggest that vertebrate homeodomain proteins are part of the biochemical pathway leading to the generation of the body axis.     

Monitoring of denitrification byPseudomonas aeruginosa using on-line fluorescence technique

Summary The NAD(P)H fluorescence ofPseudomonas aeruginosa dropped sharply upon addition of nitrate to an anaerobic culture, indicating that denitrification is not limited by mass transfer of nitrate through cell membrane to reach nitrate reductase. The effect of added nitrate concentration on fluorescence drop followed a typical saturation kinetics. The maximum specific denitrification rate under the studied condition was found to be 0.26±0.05 g NO ₃ ⁻ -N/g cells-hr.     

The CD3-gamma and CD3-delta subunits of the T cell antigen receptor can be expressed within distinct functional TCR/CD3 complexes.

The T cell receptor for antigen (TCR) consists of two glycoproteins containing variable regions (TCR-alpha/beta or TCR-gamma/delta) which are expressed on the cell surface in association with at least four invariant proteins (CD3-gamma, -delta, -epsilon and -zeta). CD3-gamma and CD3-delta chains are highly homologous, especially in the cytoplasmic domain. The similarity observed in their genomic organization and their proximity in the chromosome indicate that both genes arose from duplication of a single gene. Here, we provide several lines of evidence which indicate that in human and murine T cells which expressed both the CD3-gamma and CD3-delta chains on their surface, the TCR/CD3 complex consisted of a mixture of alpha beta gamma epsilon zeta and alpha beta delta epsilon zeta complexes rather than a single alpha beta gamma delta epsilon zeta complex. First, a CD3-gamma specific antibody failed to co-immunoprecipitate CD3-delta and conversely, several CD3-delta specific antibodies did not coprecipitate CD3-gamma. Secondly, analysis of a panel of human and murine T cell lines demonstrated that CD3-gamma and CD3-delta were expressed at highly variable ratios on their surface. This suggested that these chains were not expressed as a single complex. Thirdly, CD3-gamma and CD3-delta competed for binding to CD3-epsilon in transfected COS cells, suggesting that CD3-gamma and CD3-delta formed mutually exclusive complexes. The existence of these two forms of TCR/CD3 complexes could have important implications in the understanding of T cell receptor function and its role in T cell development.     

Effects of microorganisms on effective oxygen diffusion coefficients and solubilities in fermentation media

Effective oxygen diffusion coefficients and solubilities were measured for submerged cultures of Saccharomyces cerevisiae, Escherichia coli, and Penicillium chrysogenum. Both effective oxygen diffusion coefficients and solubilities were found to decrease with increasing cell concentrations in the fermentation media. Comparison of the experimental results of effective oxygen diffusion coefficients in fermentation media with values theoretically predicted on the assumption of unpenetrable microbial cells indicates that oxygen molecules diffuse through the cells during the diffusion process. Within the cell concentration range of typical submerged fermentations, the effective oxygen diffusion coefficient of the fermentation media can be described as D(e) = A(1)f + A(2)f(2). In this equation, fis the cell volume fraction and both A(1) and A(2) are functions of the shape of the cells and the ratio of effective oxygen diffusion coefficient in microbial cells to that in the medium.     

Pregnancy diagnosis by the ultrasonographical device and observation of fetal growth in the squirrel monkey (Saimiri sciureus)

This paper describes usefulness of the ultrasonographical device (USD) for the diagnosis of pregnancy and the observation of fetal growth in the squirrel monkeys (Saimiri sciureus) conceived under group breeding conditions. Pregnancy was diagnosed on the basis of the detection of gestational sac (GS) in the uterus. The GS was first detected 127 +/- 10 days before delivery. The heart beat of embryo was detected around 114 days before delivery. It was able to judge conditions of fetal growth by measuring the size of GS and the biparietal diameter. No difference in uterine size between nonpregnant and pregnant animals was observed 135 days before delivery.     

Cytolytic activity of Ia-restricted T cell clones and hybridomas: evidence for a cytolytic mechanism independent of interferon-gamma, lymphotoxin, and tumor necrosis factor-alpha

Ten different helper T cell (Th) hybridomas that are specific to Ia or antigen plus Ia were found to express nonspecific cytolytic activity toward the cytotoxin (CT)-resistant P815 cells upon activation with either Con A or a monoclonal anti-T3 antibody (T3-mAb). In contrast to cytolytic Th1 clones which secrete high levels of interferon-gamma (IFN-gamma) and cytotoxin (CT) (lymphotoxin (LT, also known as TNF-beta) or tumor necrosis factor-alpha (TNF-alpha], these Th hybridomas produce low or undetectable levels of IFN-gamma and CT. No inhibitory activity of IFN-gamma and CT was observed in culture supernatants of activated Th hybridomas. Double-chamber experiments demonstrated that CT-sensitive L929 cells when physically separated from activated Th1 clones were killed by membrane-permeable CT. Under identical experimental conditions, lysis of P815 cells did not occur. Moreover, activation of Th hybridomas directly in wells containing the CT-sensitive L929 cells failed to induce target cell lysis. This confirms that these Th hybridomas produce little CT and argues against high local concentrations of CT being responsible for Th hybridoma-mediated killing of P815 cells. Finally, a polyclonal rabbit antiserum to rTNF-alpha, which strongly and specifically inhibited CT-mediated and Th1 clone-mediated killing of L929 cells, failed to inhibit P815 lysis by activated Th1 clones and Th hybridomas. These observations establish that a cytolytic mechanism independent of IFN-gamma, LT, and TNF-alpha is responsible for lysis of CT-resistant target cells.     

Internal structure of the postcapillary high-endothelial venules of rodent lymph nodes and Peyer's patches and the transendothelial lymphocyte passage

Scanning electron microscopy (SEM) shows that the postcapillary high-endothelial venules of lymph nodes and Peyer's patches consist of two segments each with a different surface relief: a proximal segment with a cobblestone surface pattern and a distal segment of interlacing cytoplasmic plates. Both segments have deep adluminal crevices in which lymphocytes are lodged. The internal structural configuration of this endothelium has been examined by transmission electron microscopy (TEM) of serial sections of lymph nodes and Peyer's patches of mice, rats, and guinea pigs. The serial sections revealed that the endothelial cell bodies and their cytoplasmic extensions were disposed in a direction generally lateral to the luminal surface and intruded into the intercellular spaces of similarly disposed neighboring endothelial cells, resulting in a complex interlacing cellular pattern. Lymphocytes penetrated the endothelial cell body and secondarily followed an intracellular pathway through which they entered the extravascular compartment. At the exposed surfaces of the adluminal venule wall, recirculating lymphocytes were seen in SEM images to enter the endothelium by penetrating the endothelial cell body. The mode of migration of lymphocytes lodged in the endothelial crevices could be determined by SEM and has been examined by TEM of serial sections. At these locations as at the exposed surfaces, lymphocytes also entered the venule by penetrating the endothelial cell body. At both sites this transcellular pathway was followed by lymphocyte entry into the intercellular spaces from which they migrated into the extravascular compartment.