Identification of a high molecular weight macrophage colony-stimulating factor as a glycosaminoglycan-containing species.

Chinese hamster ovary cells transfected with a 4.0-kilobase macrophage colony-stimulating factor (M-CSF) cDNA express two different M-CSF species; one has an apparent molecular weight of 85,000 and is identified as a homodimer of a 43-kDa subunit, and the other has an indeterminate structure greater than 200 kDa. In this study, we investigated the structure of the high molecular weight M-CSF by immunochemical procedures. The high molecular weight M-CSF was easily purified, since it bound tightly to DEAE-Sephacel and eluted at a characteristically high salt concentration. The high molecular weight M-CSF migrated as a diffuse band of over than 200,000 on nonreducing sodium dodecyl sulfate-polyacrylamide gels. Analysis of the same samples under reducing conditions revealed that the larger species consisted of a heteromer of the 43- and 150-200-kDa M-CSF subunits. Digestion of the 150-200-kDa M-CSF subunit with chondroitinase, which degrades the chondroitin sulfate glycosaminoglycan chain, yielded a 100 kDa band. This species was secreted instead of 150-200-kDa species when the cells were cultured in the presence of beta-D-xyloside, which inhibits the elongation of the chondroitin sulfate glycosaminoglycan chain in proteoglycans, providing additional evidence for the existence of a chondroitin sulfate chain in the 150-200-kDa M-CSF subunit. Removal of O- and N-linked carbohydrate from the 150-200-kDa subunit yielded a polypeptide chain with a larger molecular mass (approximately 45 kDa) than that of the 43-kDa subunit (approximately 25 kDa). Collectively, these results indicate that the 150-200-kDa M-CSF subunit is a proteoglycan with a core protein that may be an alternatively processed form of M-CSF.     

Regulatory mechanism of reagin production in mice at the T cell level. I. Suggestive evidence for the generation of class specific PPD-reactive helper T cell population in Mycobacterium-primed cells.

Helper T cell activities specific for purified protein derivative (PPD) generated by immunization with Mycobacterium tuberculosis (Tbc) or PPD were investigated concerning adoptive IgE and IgG antibody responses. It is interesting that preferential triggering activity of IgG antibody response was observed when PPD-reactive cells from mice immunized with Tbc were used as a helper cell source. The selective triggering of IgG B cells by Tbc-primed cells was consistently observed using DNP-primed B cell populations from mice immunized with DNP-carrier conjugate in either ICFA or alum. T cell dependency of helper activity was demonstrated by the fact that treatment of Tbc-primed cells with anti-Thy 1 antiserum plus complement abolished their helper activity. We also demonstrated that purified T cell populations selectively triggerred IgG B cells. Selective triggering of IgG B lymphocytes by Tbc-primed T cells may not be due to the influence of suppressor T cells supposedly present in Tbc-primed cells since this selectivity was not affected by X-irradiation of Tbc-primed T cell populations which may inactivate suppressor T cells. Furthermore, passive transfer of Tbc-primed cells into normal recipient mice, the condition which may detect the suppressor T cell effect much more sensitively in IgE production, or preimmunization with Tbc 2 weeks before, did not suppress primary anti-DNP IgE antibody response to DNP-PPD. Thus, the observations presented here are favorable to the concept of the presence of IgG class-specific helper T lymphocytes. Furthermore, PPD-reactive T cells from mice immunized with PPD itself exerted their helper function for triggering B cells of both IgE and IgG classes. This may also indicate that some of the components associated with Tbc other than PPD might negatively affect the development of PPD-reactive helper T cells specific to the IgE class. The generation of such IgG-specific T cell activity in the presence of Tbc will be discussed in the light of the T cell population involved in the regulation of antibody responses of different immunoglobulin classes.     

Indoor environmental study and post occupancy evaluation in an office environment

1. 1. The objective of this paper is to investigate the indoor environment from the viewpoint of interaction between physical environment and the human responses. The field survey has been conducted over 1 year.

2. 2. A continuous measurement has been carried out for 1 week and distribution of variables have been measured for 1 day.

3. 3. The attitude of workers was investigated by a questionnaire.

4. 4. As the result, average luminance represented more than 1000 lx in the new building, in contrast with less tha 300 lx in the existing building.

5. 5. There was a significant difference of the occupants' response to the light environment between the two buildings.

6. 6. Measured thermal conditions are on the edge of the ASHRAE comfort envelope in summer, and in the neighborhood of the lower dry limit of the envelope in spring.

7. 7. The occupants' evaluations were remarkably changed before and after the moving. The office environment is better than that of the factory.

Studies on the Protection Test of Pertussis Vaccine. Immunization Period and Protectivity

In order to confirm the data reported in the previous papers, variously prepared pertussis vaccines were employed in the present investigation. Pertussis organisms grown either on a solid or in a liquid semisynthetic medium were treated by: (1) heating at 56 C for 30 min, (2) storage in 0.1% formalin at 37 C for 5 days, (3) storage in 0.1% formalin at 25 C for 5 days, and (4) simple addition of sodium ethyl-mercuri thiosalicylate (merthiolate) as a preservative. Freeze-dried vaccines were made from these preparations four and a half months later. Mice were immunized intraperitoneally and challenged intracerebrally with a virulent strain of Bordetella pertussis 10 or 17 days later. The data were analyzed statistically assuming the probit corresponding to the percentage of survivors at any dose be a linear function of the logarithms of the dose. The 50% effective doses (ED₅₀) and slopes of each vaccine were found to be uniform. More accurate estimates of ED₅₀ were obtained by employing a pooled slope in each experiment. From these ED₅₀ values, the relative potency was estimated by comparing the value of a vaccine to that of a dried merthiolate-vaccine. For vaccines derived from solid cultures, with an immunization period of 17 days, the relative potency of the vaccine heated at 56 C was 0.63 (95% fiducial limits=0.52 to 0.76); the value for the formalinized vaccine at 37 C was 0.40 (0.30 to 0.53) and one at 25 C was 0.51 (0.34 to 0.77). Vaccines derived from liquid cultures showed a relative potency of 20 to 50% less than that of corresponding vaccine derived from a solid culture. The potency obtained for the 17 day immunization period was usually higher than that for the 10 day period. Using the overall-pooled slope, an experimental design which will be appropriate for statistical analysis is discussed.     

Impact of Pulsed Nd:YAG Laser Irradiation on the Growth and Mortality of the Biofilm Forming Marine Bacterium Pseudoalteromonas carrageenovora

The impact of pulsed laser irradiation on the marine biofilm forming bacterium Pseudoalteromonas carrageenovora was investigated in the laboratory by monitoring mortality and the post-irradiation growth pattern. The impact of laser irradiation on bacterial mortality increased with the duration of irradiation. Laser irradiation at 532 nm (0.1 J cm m 2 ) for 15 min resulted in a 53% cell mortality immediately after irradiation. However, the impact after a period of 5 h (delayed impact) was more severe. The growth pattern of irradiated samples showed a prolonged lag phase compared to the reference, due to a reduction in total viable counts (TVC) in the irradiated samples. Nucleic acid staining is suggested to be a promising technique for monitoring laser inflicted bacterial mortality. Thus, the results suggest that laser irradiation could be considered as an alternative technique to reduce the number of biofilm forming bacteria and thereby biofilm formation on hard surfaces.     

Ability of Three Kind of Imidazole Dipeptides,Carnosine, Anserine,and Balenine,to Interact with Unsaturated Fatty Acid-Derived Aldehydes and Carbohydrate-Derived Aldehydes

International Journal of Peptide Research and Therapeutics - Imidazole dipeptides (IDPs) such as carnosine (CAR), anserine (ANS), and balenine (BAL) are widely distributed in the skeletal muscle of...     

Synthesis of peptide conjugates with vitamins for induction of antigen‐specific immunotolerance

In this report, we designed conjugates of an antigen peptide with the immunosuppressive vitamins all‐trans retinoic acid (ATRA) and vitamin D3 for efficient induction of antigen‐specific immunotolerance. We established a synthetic scheme for the preparation of the peptide‐vitamin conjugates, which the chemically unstable vitamins tolerated. Among the obtained conjugates, the ATRA conjugate successfully suppressed inflammatory effects in macrophages and dendritic cells and induced antigen presentation in dendritic cells. This synthetic method of conjugate is conceivably applicable to other antigen peptides for induction of antigen‐specific immunotolerance.     

Spine and dine: A key defensive trait promotes ecological success in spiny ants

A key focus of ecologists is explaining the origin and maintenance of morphological diversity and its association with ecological success. We investigate potential benefits and costs of a common and varied morphological trait, cuticular spines, for foraging behavior, interspecific competition, and predator–prey interactions in naturally co‐occurring spiny ants (Hymenoptera: Formicidae: Polyrhachis) in an experimental setting. We expect that a defensive trait like spines might be associated with more conspicuous foraging, a greater number of workers sent out to forage, and potentially increased competitive ability. Alternatively, consistent with the ecological trade‐off hypothesis, we expect that investment in spines for antipredator defense might be negatively correlated with these other ecological traits. We find little evidence for any costs to ecological traits, instead finding that species with longer spines either outperform or do not differ from species with shorter spines for all tested metrics, including resource discovery rate and foraging effort as well as competitive ability and antipredator defense. Spines appear to confer broad antipredator benefits and serve as a form of defense with undetectable costs to key ecological abilities like resource foraging and competitive ability, providing an explanation for both the ecological success of the study genus and the large number of evolutionary origins of this trait across all ants. This study also provides a rare quantitative empirical test of ecological effects related to a morphological trait in ants.     

Recent progress of animal transplantation studies for treating articular cartilage damage using pluripotent stem cells

Focal articular cartilage damage can eventually lead to the onset of osteoarthritis with degradation around healthy articular cartilage. Currently, there are no drugs available that effectively repair articular cartilage damage. Several surgical techniques exist and are expected to prevent progression to osteoarthritis, but they do not offer a long‐term clinical solution. Recently, regenerative medicine approaches using human pluripotent stem cells (PSCs) have gained attention as new cell sources for therapeutic products. To translate PSCs to clinical application, appropriate cultures that produce large amounts of chondrocytes and hyaline cartilage are needed. So too are assays for the safety and efficacy of the cellular materials in preclinical studies including animal transplantation models. To confirm safety and efficacy, transplantation into the subcutaneous space and articular cartilage defects have been performed in animal models. All but one study we reviewed that transplanted PSC‐derived cellular products into articular cartilage defects found safe and effective recovery. However, for most of those studies, the quality of the PSCs was not verified, and the evaluations were done with small animals over short observation periods. Large animals and longer observation times are preferred. We will discuss the recent progress and future direction of the animal transplantation studies for the treatment of focal articular cartilage damages using PSCs.