Formation of the junctions between lymph follicles in the Peyer's patches even before postweaning activation.

Peyer's patches (PPs), which contain an abundance of B and T cells, play a key role in inducing pivotal immune responses in the intestinal tract. PPs are defined as aggregated lymph follicles, which consist of multiple lymph follicles (LFs) that may interact with each other in a synergistic manner. LFs are thought to be spherical in shape; however, the characteristics of their structure are not fully understood. To elucidate changes in the structure of PPs as individuals grow, we generated serial 2D sections from entire PPs harvested from mice at 2, 4, and 10 weeks of age and performed a 3D analysis using a software, Amira. Although the number of LFs in PPs was not changed throughout the experiment, the volume and surface area of LFs increased significantly, indicating that LFs in PPs develop continuously by recruiting immune cells, even after weaning. In response to the dramatic changes in the intestinal environment after weaning, the development of germinal centers (GCs) in LFs was observed at 4 and 10 weeks (but not 2 weeks) of age. In addition, GCs gradually began to form away from the center of LFs and close to the muscle layer where export lymphatic vessels develop. Importantly, each LF was joined to the adjacent LF; this feature was observed even in preweaning nonactivated PPs. These results suggest that PPs may have a unique organization and structure that enhance immune functions, allowing cells in LFs to have free access to adjacent LFs and egress smoothly from PPs to the periphery upon stimulation after weaning.

Effects of daily management changes on behavioral patterns of a solitary female African elephant (Loxodonta africana) in a zoo.

This study investigated the effects of changes in daily management on behavior of a solitary female elephant in a zoo. The activity budget and space utilization of the subject and the management changes were recorded for 1 year after the conspecific male died. The observation days could be categorized into five clusters (C1-C5) by the characteristic behavioral pattern of each day. C1 had the highest percentage of resting of all clusters, and was observed after the loss of the conspecific and the beginning of use of the indoor exhibition room at night. C2, which had the highest percentage of stereotypy of any cluster, was observed after the beginning of habituation to the indoor exhibition room. Also, when the time schedule of management was changed irregularly, the subject frequently exhibited stereotypic pacing (C2, C4). The subject tended to rest when exhibiting lameness in the left hind limb (C3). In C5, activity reached a high level when she could utilize a familiar place under a stable management schedule. These results indicate that management changes affected the mental stability of an elephant in the early stage of social isolation.

Evaluation of newly developed oxygen meters with multi-channels and disposable oxygen electrode sensors for antimicrobial susceptibility testing.

The objective of this study was to investigate the applicability of new oxygen meters with multi-channels and disposable oxygen electrode sensors (DOX-96) on the antimicrobial susceptibility testing of clinical bacterial isolates. The oxygen amount in the wells of 96-well plates was converted into current through electrodes. Bacterial inoculation decreased the oxygen amount in the wells because viable bacteria consume the oxygen. On the other hand, a failure of bacteria to consume oxygen was observed in the presence of potent antimicrobial agents, representing a serious arrest of bacterial metabolism usually leading to stasis or death. Based on these results, the minimum inhibitory concentration was determined by DOX-96 (MICdox). The MICdox showed good agreement with MIC measured by the standard broth microdilution method (98.2%). DOX-96 was also useful for turbid samples such as Mueller-Hinton broth containing 0.1% lipid emulsion. The MICdox in turbid samples showed good agreement with those in clear samples (100.0%). These results indicate that the newly developed DOX-96 is very useful in antimicrobial susceptibility testing even in turbid clinical samples such as colloidal products and turbid biological components.

Antimicrobial activity of saturated fatty acids and fatty amines against methicillin-resistant Staphylococcus aureus.

The objective of this study was to investigate the antimicrobial activities of saturated fatty acids and fatty amines against methicillin-resistant Staphylococcus aureus (MRSA). The antimicrobial activity of saturated fatty acids and fatty amines was determined by oxygen meters with multi-channels and disposable oxygen electrode sensors (DOX-96). Lauric acid, the most effective among the saturated fatty acids, showed antimicrobial activity at 400 microg/ml against methicillin-susceptible Staphylococcus aureus (MSSA) and MRSA. The minimal inhibitory concentration (MIC) of fatty amines depended on each hydrophobic chain length. The MIC of myristylamine was 1.56 microg/ml; most effective of the fatty amines. In time-kill curves, lauric acid and myristylamine produced a bactericidal effect and a bacteriostatic effect at 4-fold the MIC, respectively. The antimicrobial activities of lauric acid and myristylamine were decreased by human plasma. Cytotoxicity of 3 saturated fatty acids and 3 fatty amines was examined in cultured endothelial cells. Although cytotoxicity of fatty amines was severer than that of saturated fatty acids, myristylamine showed the highest value of apparent therapeutic index among them. DOX-96 was useful for screening antimicrobial substances, especially in the case of insoluble substances. We found that myristylamine showed anti-MRSA activity comparable to that of vancomycin and teicoplanin.