Immune synergistic oligodeoxynucleotide from Lactobacillus rhamnosus GG enhances the immune response upon co‐stimulation by bacterial and fungal cell wall components

Bacterial genomic DNA has recently been shown to elicit a highly evolved immune defense. This response can be selectively triggered for a wide range of therapeutic applications, including use as a vaccine adjuvant to immunotherapies for allergy, cancer, and infectious diseases. Previously, we identified a low‐concentration immune synergistic oligodeoxynucleotide (iSN‐ODN, named iSN34) from Lactobacillus rhamnosus GG that has immunosynergistic activity upon costimulation of target cells with ligands of Toll‐like receptor 9 (TLR9). Here, we extend that observation by demonstrating the synergistic induction (in mouse splenocytes) of IL‐6 by the combination of iSN34 with cell wall components of bacteria and fungi. We observed that splenocytes pretreated with iSN34 and then costimulated with agonists for TLR1/2 (Pam₃CSK₄), TLR4 (lipopolysaccharide), or TLR2/6 (Zymosan) exhibited enhanced accumulation of IL‐6. These results suggested that the combination of iSN34 with TLR1/2, TLR4, or TLR2/6 agonists may permit the induction of a potent immune response.     

Physical and chemical characteristics of the blackened portion of Japanese persimmon (Diospyros kaki)

The heartwood of the Japanese persimmon tree (Diospyros kaki) becomes black on rare occasions and has been highly esteemed as a substitute for ebony. We attempted to clarify how the physical, mechanical, chemical, and biodegradation properties differ between sapwood and blackened heartwood. The specific gravity, equilibrium moisture content, modulus of rupture, and modulus of elasticity in the blackened heartwood were higher, and the loss tangent was lower, than those in sapwood. Furthermore, the blackened portion was more resistant to fungal and termite attacks. A section of heartwood was dark-brown, and the specific gravity and mechanical properties of this portion were slightly lower than those in sapwood. The dark-brown portion was speculated to be a sign or interrupted state of fungal attack. The blackening substance was bound tightly to cell wall components and could not be extracted with any of the organic solvents used. The findings of trace element analysis using inductively coupled plasma-mass spectrometry showed that the boron content was markedly high in the blackened portion. The findings obtained here suggest a role of boron in the antifungal properties and the blackening phenomenon of Japanese persimmon.Part of this work was presented at the 50th Annual Meeting of the Japan Wood Research Society, Kyoto, April 2000     

Glycation and phosphorylation of beta-lactoglobulin by dry-heating: effect on protein structure and some properties

Beta-lactoglobulin (beta-Lg) was glycated with maltopentaose and subsequently phosphorylated by dry-heating in the presence of pyrophosphate to investigate the structural and functional properties of phosphorylated beta-Lg. The circular dichroism spectra showed that the change of the secondary structure in the beta-Lg molecule by glycation and subsequent phosphorylation was small. The differential scanning calorimetry thermograms of beta-Lg showed that the denaturation temperature of the most stable domain was only slightly affected, whereas the retinol-binding activity of beta-Lg was somewhat reduced by glycation and subsequent phosphorylation. These results indicated that the conformational changes of the beta-Lg molecule by glycation and subsequent phosphorylation were mild. The anti-beta-Lg antibody response was somewhat reduced by glycation, but significant changes were not observed by phosphorylation. Although the stability of beta-Lg against heat-induced insolubility was improved by glycation alone, it was further enhanced by phosphorylation. The calcium phosphate solubilizing ability of beta-Lg was enhanced by phosphorylation following glycation.     

Juvenile bird acanthocephalans recovered incidentally from raccoon dogs (Nyctereutes procyonoides viverrinus) on Yakushima Island, Japan

Helminth parasites were collected from 14 raccoon dogs (Nyctereutes procyonoides viverrinus) and 2 Japanese weasels (Mustela itatsi sho) on Yakushima Island, Kagoshima Prefecture (Kyushu), Japan. In addition to trematodes and nematodes reported separately, everted cystacanths of 3 bird acanthocephalans (Mediorhynchus robustus in 5 animals, Porrorchis leibyi in 2 animals, and Sphaerirostris lanceoides in 1 animal) were collected from the raccoon dogs introduced recently on this World Natural Heritage Area. A cystacanth of P. leibyi was collected also from a weasel indigenous to this island that were killed by traffic accidents. Recovery of everted cystacanths of M. robustus from the intestine of raccoon dogs was remarkably frequent (36%), suggesting that this acanthocephalan should be highly prevalent in passerine hosts on Yakushima Island, and the animals might vigorously take insect hosts or paratenic hosts. Furthermore, the present report is the second on M. robustus after its first record in Japan (Aichi Prefecture, Honshu) as M. garruli from a Eurasian jay (Garrulus glandarius japonicus) by Yamaguti in 1939. Porrorchis leibyi has never been recorded in Japan.     

Effect of bovine lactoferrin on the immune responses of captive bottlenosed dolphins (Tursiops truncatus) being transported over long distances

Bovine lactoferrin was administered orally, in feed, to six bottlenosed dolphins (Tursiops truncatus) before they were transported for approximately six hours; their stress responses were compared with those of five untreated dolphins. During the journey the dolphins had an increased plasma concentration of cortisol, and lymphopenia, eosinopenia and mild neutrophilia, indicating a stress response. The administration of lactoferrin did not affect the function of the dolphins' polymorphonuclear leucocytes, but affected their leucogram by maintaining the number of circulating eosinophils.     

Development and biological properties of a new live attenuated mumps vaccine

To develop a new live attenuated mumps vaccine, a wild mumps Y7 strain isolated from a patient who developed mild parotitis was treated with nitrosoguanidine and ultraviolet, followed by selection of a temperature-sensitive clone. The selected clone, Y125, showed stable temperature-sensitivity in Vero cells. Intraspinal inoculation of marmosets with the Y125 produced only minimal histopathological changes, while intracerebral inoculation of neonatal rats revealed that the Y125 did not cause hydrocephalus. Both these effects of the Y125 were similar to those of the non-neurovirulent Jeryl Lynn strain. Furthermore, subcutaneous inoculation of the Y125 induced high levels of neutralizing antibodies in all Cercopithecus monkeys examined. Although the safety and immunogenicity should be confirmed in further field trials in humans, the present results indicate that the Y125 could be a promising vaccine candidate.     

Relationship between pH and Temperature in the Ruminal Fluid of Cows, Based on a Radio-Transmission pH-Measurement System

To assess the relationship between pH and temperature in the ruminal bottom fluid, circadian changes were monitored using cows fed a control diet (C diet) or a rumen acidosis-inducing diet (RAI diet) by using a wireless radio-transmission pH- measurement system. These two parameters were measured simultaneously at 10-min intervals on day 14 after commencement of feeding. Compared to the mean ruminal pH for 60 min immediately after the morning feeding (0 hr), significantly lower pH was noted 3-13 hr later (P<0.05) and 4-19 hr later (P<0.01) in cows fed the C and RAI diets, respectively, although the reduction in the latter was much higher than that in the former. In contrast, significantly higher ruminal temperature was found at 8 and 12-14 hr later (P<0.05) and 6, 8, and 10-19 hr later (P<0.01) in cows fed the C and RAI diets, respectively. A significant negative correlation was observed between the lowest ruminal pH and its corresponding ruminal temperature in cows fed the C and RAI diets (r=-0.722 and -0.650, P<0.01, respectively), suggesting active fermentation and volatile fatty acid production in the rumen. However, ruminal pH profiles may not be predictable by measuring only ruminal temperature because decreases in ruminal pH were preceded by increases in ruminal temperature, and circadian changes in pH and temperature were associated with ruminal fermentation.