A Hodgkin's disease cell line, KM-H2, shows biphenotypic features of dendritic cells and B cells.

The origin of Reed-Sternberg (RS) cells, the neoplastic cells of Hodgkin's disease, has long remained controversial. Dendritic cells (DCs) are highly specialized antigen-presenting cells that have the unique capacity to prime naive T cells, and they may be progenitors of RS cells in a population of Hodgkin's disease cells. In this study, the KM-H2 cell line, previously established from a patient with Hodgkin's disease of mixed cellularity, was reevaluated for its cellular derivation, particularly in terms of DCs. KM-H2 cells were demonstrated to carry the newly proposed DC-associated molecules fascin, CD83, and DEC-205, as well as costimulatory molecules such as CD40, CD80, and CD86. In addition, KM-H2 cells were shown to be able to potently stimulate peripheral blood T cells and to have the strong endocytotic activity of fluorescein isothiocyanate-dextran. On the other hand, KM-H2 cells were shown to have variable-diversity-joining recombination of the immunoglobulin H gene, although they did not express any subclasses of immunoglobulin and they were negative for CD79a and CD79b. In addition, KM-H2 cells produced the messenger RNA of the Pax-5 gene. These findings lead to a hypothesis that KM-H2 cells originated from the cells that had differentiated through the possible common DC-B-cell progenitors along the newly proposed pathway.

Molecular cloning, enhancement of expression efficiency and site-directed mutagenesis of rat epidermal cystatin A.

A rat cystatin A cDNA clone was isolated from a lambda ZAP library representing newborn rat skin mRNA by screening with a synthetic oligonucleotide designed from amino acid sequence 15-23 of the cysteine proteinase inhibitor. The obtained clone contained a partial coding region of the inhibitor, lacking the 5'-untranslated region and coding sequence for the NH(2)-terminal 13 residues. The amino acid sequence deduced from the base sequence, Glu14-Phe103, coincided with that determined at the amino acid level. To obtain the recombinant cystatin A protein, the DNA was fused with a synthetic linker encoding its missing N-terminal 17 residues and introduced into an expression vector, pMK2. In Escherichia coli, however, the expression level of the semi-synthetic gene was low, 0. 5 mg of the purified recombinant protein per 1 liter culture being produced. Changing of the codon usage of the N-terminal region in a pET-15b expression system led to an increase in the yield depending on the instability of the putative secondary structure around an initiation codon of the mRNA. The expressed cystatin A showed identical characteristics with the authentic form except for the absence of the N-terminal acetyl blocking group. Using the expression system, two kinds of point mutation, the conservative Val54 in the first loop QxVxG region being changed to Lys and Glu, were introduced, but there was almost no effect on the inhibitory activity toward papain. This suggests that the conserved Val in the reactive site is not restricted and that the hydrophobicity of the position is not essential for the activity of rat cystatin A.

[Subjective well-being differences in disabled women as related to length of bedridden period].

A cross-sectional study of disabled women living at home, aged 65 years and more, was conducted in Osaka-city to examine difference in subjective well-being in two groups differing in length of bedridden period, and to elucidate correlates of subjective well-being. A short-period group was composed of 43 disabled women whose bedridden periods were shorter than two years, and a long-period group of 63 women was randomly selected from disabled women of the corresponding age having a bedridden experience of five or more years. The two groups were interviewed by public health nurses using the Philadelphia Geriatric Center Morale Scale. The subjective well-being was lower in the short-period group than in the long-period group. This result indicates the necessity to provide adequate support to the disabled elderly early on, not only from the physical stand point but also for consideration of subjective well-being. Other correlates of subjective well-being in the disabled women were age, number of family members and caregivers, economic state, pain, ophthalmopathy and utilization of domiciliary care services. Existence of a plan for home visits by public health nurses was strongly related to subjective well-being, possibly indicating that the visiting plan was made with some consideration of subjective well-being. These need to be further development of adequate support systems that consider the subjective well-being of the disabled elderly.