Cancer chemotherapy and somatic cell mutation

The occurrence of a second neoplasm is one of the major obstacles in cancer chemotherapy. The elucidation of the genotoxic effects induced by anti-cancer drugs is considered to be helpful in identifying the degree of cancer risk. Numerous investigations on cancer patients after chemotherapy have demonstrated: (i) an increase in the in vivo somatic cell mutant frequency (Mf) at three genetic loci, including hypoxanthine–guanine phosphoribosyl-transferase (hprt), glycophorin A (GPA), and the T-cell receptor (TCR), and (ii) alterations in the mutational spectra of hprt mutants. However, the time required for and the degree of such changes are quite variable among patients even if they have received the same chemotherapy, suggesting the existence of underlying genetic factor(s). Accordingly, some cancer patients prior to chemotherapy as well as patients with cancer-prone syndrome have been found to show an elevated Mf. Based on the information obtained from somatic cell mutation assays, an individualized chemotherapy should be considered in order to minimize the risk of a second neoplasm.     

Further characterization of a conserved actin-binding 27-kDa fragment of actinogelin and alpha-actinins and mapping of their binding sites on the actin molecule by chemical cross-linking

A conserved actin-binding domain (Mr = 27,000) of rat hepatic actinogelin, rat skeletal muscle, and chicken gizzard alpha-actinins (Mimura, N., and Asano, A. (1986) J. Biol. Chem. 261, 10680-10687) was separated into two components having different isoelectric points (peptides A and B) by chromatofocusing. Thermolysin digestion of peptide A generated peptide B with concomitant loss of peptide A. Amino acid compositions and tryptic maps of peptides A and B also demonstrated that peptide A is a precursor of peptide B upon thermolysin digestion. All of peptides A and B retained the activity to bind with F-actin competitively to each other. By the gel-filtration method, it was also shown that the native actin-binding 27-kDa fragments are monomeric and globular. The non-actin-binding 50- or 53.5-kDa fragment of actinogelin/alpha-actinins was, however, found to be asymmetric and dimeric in the native state. Chemical cross-linking of the 27-kDa fragment with F-actin with a water-soluble carbodiimide produced at least four different complexes (I-IV). Chemical cleaving analysis of the cross-linked products (complexes I and II) indicated that the 27-kDa fragment possesses two possible binding sites on actin at the NH2-terminal residues 1-12 (for complex I) and at residues spanning 86-119 or 123 (for complex II).     

A novel MHC class I-related molecule expressed on mouse thymic stroma cells and mature lymphocytes

A monoclonal antibody (mAb) TP-3 has been established by immunizing rats with the BALB/c mouse thymic epithelial cell line TEL-2. The TP-3 antigen is expressed on stroma cells of thymus, spleen, and lymph node in syngeneic BALB/c mice (H-2 ^d ). This antigen is also expressed at a low level on the cell surface of immature thymocytes, and at a high level on mature T and B cells. In allogeneic mice such as C57BL/6 (H-2 ^b ) or C3H (H-2 ^k ), no cells expressed the TP-3 antigen. Using H-2 congenic mice, reactivity with mAb TP-3 was found to map to a region of H-2D ^d L ^d or between D ^d and Qa, suggesting that TP-3 is a major histocompatibility complex (MHC) class I antigen. However, immunoprecipitation analysis indicated that this antigen is not identical to the classical mouse class I molecules in terms of molecular size, antigenicity, and tissue distribution.     

Gene structure and expression of a tobacco endochitinase gene in suspension-cultured tobacco cells

We have isolated and characterized the genomic clone CHN50 corresponding to tobacco basic endochitinase (E.C.3.2.1.14). DNA sequence and blotting analysis reveal that the coding sequence of the gene present on CHN50 is identical to that of the cDNA clone pCHN50 and, moreover, the CHN50 gene has its origin in the progenitor of tobacco, Nicotiana sylvestris. Tobacco basic chitinases are encoded by a small gene family that consists of at least two members, the CHN50 gene and a closely related CHN17 gene which was characterized previously. By northern blot analysis, it is shown that the CHN50 gene is highly expressed in suspension-cultured tobacco cells and the mRNA accumulates at late logarithmic growth phase. To identify cis-DNA elements involved in the expression of the CHN50 gene in suspensioncultured cells, the chimeric gene consisting of 1.1 kb CHN50 5 upstream region fused to the coding sequence of -glucuronidase (GUS) was introduced by electroporation into protoplasts isolated from suspension-cultured tobacco cells. Transient GUS activity was found to be dependent on the growth phase of the cultured cells, from which protoplasts had been prepared. Functional analysis of 5 deletions suggests that the distal region between -788 and -345 contains sequences that potentiate the high-level expression in tobacco protoplasts and the region (-68 to -47) proximal to the TATA box functions as a putative silencer.     

Identification of (Na,K)ATPase inhibitor in brine shrimp,Artemia salina,as long-chain fatty acids

Summary An inhibitory activity to (Na,K)ATPase was found in cell extracts of the brine shrimp, Artemia salina, irrespective of its developmental stages. Organic solvent extraction together with gas chromatographic analysis reveals that the inhibitory activity is due to long-chain, non-esterified fatty acids and their derivatives. Unsaturated fatty acids, especially with cis-configuration, are more effective in inhibition than saturated ones.Abbreviations ATPase adenosine triphosphatase - EDTA ethylenediamine-tetraacetate - TLC thin-layer chromatography     

A new stichaeid fish of the genusAlectrias from Mutsu Bay,northern Japan

Alectrias mutsuensis sp. nov. is described on specimens collected from Mutsu Bay, Aomori Pref. This new species is rather deep dweller compared to other species of the genus, living on muddy bottoms, about 20 to 40 meters deep, and is diagnosed by the following characters: dermal crest on head reaching to occipital region; distance between end of dermal crest and dorsal fin origin less than eye diameter; end of anal fin separated from caudal fin; anterior dorsal spines before anal origin, slender and flexible; epaxial hypurals mostly fused into a single plate. A key to all nine species belonging to the subfamily Alectriinae is presented.     

Triterpenes from Periandra dulcis roots

Three oleanane triterpenes were isolated from the roots of Periandra dulcis,and identified as 3β-hydroxy-25-al-olean-18-en-30-oic acid (periandric acid I), 3β-hydroxy-25-al-olean-12-en-30-oic acid (periandric acid II) and 3-oxo-25-hydroxy-olean-12-en-30-oic acid. The former two compounds (periandric acids I and II) were identical with the aglycones obtained by hydrolysis of periandrin I and II, respectively and the latter one was a new triterpene.     

Circadian rhythms of testosterone-dependent behaviors,crowing and locomotor activity,in male Japanese quail

Summary An apparatus was devised to record crowing (mate calling by males) together with locomotor activity and recorded data was analyzed by several methods for rhythm analysis. Crowing and locomotor activity of Japanese quail held on long days were recorded during sexual development as estimated from circulating gonadotropins and testosterone. Both behaviors were testosterone-dependent but commencement of crowing preceded the increase in locomotor activity. When the two behaviors attained their maximum levels, crowing showed consistent daily rhythms in which two peaks were apparent, a major one at the onset of light and a broader one 8 hours later. Locomotor activity also showed a clear daily rhythm with a peak between the two peaks of crowing rhythm suggesting a fixed phase relationship between the two rhythms.Both rhythms free-ran under constant dim light with periods shorter than 24 h. They persisted in birds which had been castrated and then supplied with exogenous testosterone via implanted Silastic capsules. The durations of both rhythms were quite comparable to each other and they maintained a fixed phase relationship similar to that found under LD cycles.The results indicate that testosterone is essential for the induction of crowing and for the enhancement of locomotor activity but the formation of the rhythms in behavior was strictly dependent on a circadian oscillatory mechanism.Abbreviations LH luteinizing hormone - FHS follicle-stimulating hormone - LD light-dark - LDim light-dim light