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Utilizing a method called "differential display of mRNAs by means of polymerase chain reaction", the cDNA fragment of a thyroid hormone-responsive gene ZAKI-4 was cloned from cultured human skin fibroblasts. Northern blot analysis revealed that there were two ZAKI-4 mRNA species (3.4 and 1.4 kilobases (kb)), and they were up-regulated by a physiological concentration of triiodothyronine (T3). This T3 effect was abolished by the treatment with cycloheximide, indicating the possibility that gene ZAKI-4 is regulated by T3 in an indirect fashion, through an intermediate product of T3, rather directly by T3 itself. No effect of T3 on ZAKI-4 mRNA stability suggested that T3 induces the mRNA at the transcriptional level. Rapid amplification of cDNA ends confirmed the presence of two mRNA species. ZAKI-4 mRNA was detected in heart, brain, liver, and skeletal muscle but not in placenta, lung, kidney and pancreas. In skin fibroblasts and skeletal muscle, 3.4-kb mRNA was the major species, whereas 1.4-kb mRNA was dominant in heart, brain, and liver. The sequence analysis suggested that the two mRNA species arise from alternative polyadenylation and code a single protein of 192 amino acids. No homologous protein sequence was found in a data base. Elucidation of the function of ZAKI-4 gene product will provide new insights into an important role of T3 in various organs.

A novel Gram-stain-positive actinobacterium, designated RI148-Li105 T , was isolated from a lichen sample from Rishiri Island, Japan, and its taxonomic position was investigated by a polyphasic approach. 16S rRNA gene sequencing study indicated that strain RI148-Li105 T was related to the type strain of Luteimicrobium subarcticum, with a similarity of 97.8%. Cells of strain RI148-Li105 T exhibited a rod-coccus cycle. The diagnostic cell-wall diamino acid of this organism was lysine and the peptidoglycan type was found to be A4a. The predominant menaquinones were MK-8(H 2 ) and MK-9(H 2 ), and the major fatty acids were iso-C 16:0 , C 17:1 x9c and C 17:0 . The DNA G þ C content was 73.6 mol%. The major phenotypic characteristics of strain RI148-Li105 T basically corresponded to those of the genus Luteimicrobium excluding the fatty acid composition. These results suggest that strain RI148-Li105 T should be affiliated with the genus Luteimicrobium. Meanwhile, DNA-DNA hybridization and some phenotypic characteristics revealed that the strain differs from L. subarcticum. Therefore, strain RI148-Li105 T represents a novel species of the genus Luteimicrobium, for which the name Luteimicrobium album sp. nov. is proposed. The type strain of Luteimicrobium album is RI148-Li105 T ( ¼ NBRC 106348 T ¼ DSM 24866 T ).

ABSTRACT-In the majority of the models of experimental allergic rhinitis, antigen challenge has been performed by single topical instillation or perfusion with the solution. The present study was performed to establish a good model using Japanese cedar pollen, which is able to repeatedly induce allergy restricted to the upper airway. Guinea pigs sensitized with the pollen extracts were subjected to quantitative and repeated inhaling of the pollen with a devised apparatus. Following the respective challenges, the nasal cavity was washed with a new technique: Washing with physiologic saline was performed from one nostril to the other one, the latter of which was kept under slightly reduced pressure. When the animal was subjected to cedar pollen inhalation, almost all the pollens inhaled were located in the upper airway. At the 5th inhalation, nasal cavity lavage revealed that both albumin leakage and histamine release into the nasal cavity were in creased at maximum levels in 1 hr (respectively 2 mg and 3 ng/animal); and at the same time, a considerable number of leukocytes, especially eosinophils, were found migrating into the nasal cavity for at least 10 hr. The present methods can permit various analyses of allergic rhinitis and the assessment of drugs without sacrificing the animal over the long term.

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