Construction and characterization of centric circular and acentric linear chromosomes in fission yeast

Summary Gene disruption and gap repair of chromosomal DNA have been frequently employed techniques in yeast genetics. To extend the possibility of using these gene manipulations for larger genomic regions, we have examined the maximal sizes of chromosomal DNA disrupted or repaired in vivo. Here we report a simple, potentially general, method for selectively deleting a 150 kb region, or gap-filling a 100 kb region, in the fission yeast genome. This enables the generation of acentric linear chromosomes by deletion, or the cloning of large functional centromeric DNAs into circular minichromosomes by gap-filling. The fidelity of the resulting gap-filling is high, judging from partial-digestion mapping of gap-repaired DNAs. By analysing a series of such circular minichromosomes, we conclude that only a part of the repetitive centromeric region, including the central domain, is essential for mitotic and meiotic chromosome segregation. Acentric linear chromosomes, although unstable, could be maintained, indicating that it may be possible to construct an acentric vector for large DNA fragments in this organism.     

HTL V-I from Iranian Mashhadi Jews in Israel is phylogenetically related to that of Japan,India, and South America rather than to that of Africa and Melanesia

A new endemic focus of human T-lymphotropic virus type I (HTL V-I) was recently reported among Mashhadi Jews, a group of immigrants from northeastern Iran to Israel. We extracted DNAs from fresh peripheral blood mononuclear cells (PBMCs) and/or gargle mouthwash from 10 HTL V-I carriers, who consisted of members of one family, and HTL V-I-associated myelopathy (HAM) and adult T-cell leukemia (ATL) patients. Long terminal repeat (LTR) regions of proviral DNAs were sequenced and analyzed phylogenetically. In a phylogenetic tree, all the Mashhadi HTL V-I isolates belonged to subtype A, one of the three subtypes of the cosmopolitan type of HTL V-I, and made a tight cluster distinct from the other isolates of subtype A from Japan, India, the Caribbean Basin, and South America. Although a few nucleotide substitutions were observed among the clones sequenced, no characteristic sequence variation was found in different disease manifestations, even in one family or different sources of DNA preparation.     

Lymphocyte subpopulations and T-cell subsets in human oral cancer tissues: immunohistologic analysis by monoclonal antibodies

A series of T-cell-specific monoclonal antibodies (Leu-1, Leu-2a, and Leu-3a) and B-cell-specific monoclonal antibody (HLB-1) were used to detect the localization and intensity of infiltration of lymphocyte subpopulations and T-cell subsets in frozen sections of 17 patients with the oral cancer. The vast majority of the lymphocyte infiltrates in the oral cancer tissues were reactive with Leu-1. In contrast, B cells were detectable with HLB-1 in only 2 of 17 cases. Leu-2a-positive cells were dominant in four cases, whereas Leu-3a positive cells were dominant in only three cases. In seven cases, both cells infiltrated to the same degree. Leu-2a positive cells tended to be dominant in the cases with earlier clinical stages.     

Protective effects of a water-soluble extract from cultured medium of Ganoderma lucidum (Rei-shi) mycelia and Agaricus blazei murill against X-irradiation in B6C3F1 mice: Increased small intestinal crypt survival and prolongation of average time to animal death

Radioprotective effects of a water-soluble extracts from cultured medium of Ganoderma lucidum (Rei-shi) mycelia (designed as MAK) and Agaricus blazei (Agaricus) against the shortening of survival time or the injury of crypt by X-irradiation were investigated in male B6C3F1 mice. MAK and Agaricus at three different doses were mixed into basal diet into biscuits at 5, 2.5 and 1.25% and administered from 1 week before irradiation. MAK (5% group) significantly prolonged animal survival as compared with basal diet group (control group) after 7 Gy of X-ray irradiation at a dose rate of 2 Gy min(-1). At doses of 8, 10 and 12 Gy X-irradiation at a dose rate of 4 Gy min(-1) MAK (5% group) significantly increased crypt survival as compared to other groups. These results suggest that MAK can act as a radioprotective agent.     

Pathological analysis of hypertrophic cardiomyopathy simulating dilated cardiomyopathy

The pathomorphologic features of hypertrophic cardiomyopathy simulating dilated cardiomyopathy in the late stage (HCM-DCM) were compared with those of ordinary hypertrophic cardiomyopathy (HCM). Seven autopsied hearts with HCM-DCM and 11 with HCM were assessed quantitatively using an image analyzer. Unlike HCM, significant left ventricular enlargement and wall thinning were observed in HCM-DCM, and the percentage areas of massive fibrosis and disarray were significantly greater. In HCM-DCM, the disarray was distributed diffusely, whereas massive fibrosis was distributed more intensively in the ventricular septum and anterior wall than in the lateral and posterior wall. Narrowing of intramyocardial small arteries was observed more frequently in HCM-DCM, especially in the ventricular septum and anterior wall, than in HCM. These results suggest that the enlargement and wall thinning of the left ventricle in HCM-DCM are attributable to non-uniform progression of massive fibrosis, which is closely related to small-arterial lesions.     

A new nematode, Ansiruptodera scapteromi sp. nov. (Nematoda: Aspidoderidae), recovered from the Argentinean water rat Scapteromys tumidis (Waterhouse, 1837) in Uruguay

This report describes a new species of aspidoderid nematode, Ansiruptodera scapteromi sp. nov., the second species to be reported in the genus Ansiruptodera Skrjabin and Shikhobalova, 1947. The A. scapteromi sp. nov. is the first species of the genus to be recorded from a rodent host. The new species is clearly different from the only other species, A. ansirupta (Proença, 1937) Skrjabin and Shikhobalova, 1947, in that it possesses short lateral alae that terminate before the midbody; a smaller cephalic extremity; a shorter esophagus, pharynx, tail, and tail appendage; a smaller sucker; and longer spicules. The two species also differ in the numbers and arrangements of caudal papillae. A. scapteromi appears to be a parasite of capture and the water rats seem to have been infected from Edentata in Uruguay.     

Gibberellin biosynthesis in Gibberella fujikuroi: cloning and characterization of the copalyl diphosphate synthase gene

The gene coding for copalyl diphosphate synthase (CPS), which represents the first gene of the gibberellin pathway, was isolated from the rice pathogen Gibberella fujikuroi. This fungus is used commercially for the production of gibberellic acid and related gibberellins. CPS is a terpene cyclase which catalyzes the first specific step of the gibberellin (GA) pathway as it branches off from the general isoprenoid (biosynthetic) pathway at geranylgeranyl disphosphate (GGDP). A cDNA fragment of the cps gene from the fungus G. fujikuroi was amplified by RT-PCR using oligonucleotides based on amino-acid sequences which were conserved between the plant CPSs and the bifunctional CPS/KS of the fungus Phaeosphaeria sp. L487. A 588-bp fragment obtained with nested PCR was used to isolate the corresponding genomic clone of the cps gene from the wild-type λ-library. This gene consists of three exons and two introns. The three exons are 2877 bp long and encode 959 amino-acid residues. The protein shares 48% identity with the bifunctional Phaeosphaeria sp. L487 FCPS and between 16% and 18% identity to the corresponding plant CPSs. Expression of the G. fujikuroi cps gene is strongly enhanced under conditions optimized for gibberellin biosynthesis and is reduced when high amounts of ammonium are present in the medium. Gene disruption, followed by gibberellin assays and Southern-blot analysis of the transformants, demonstrated clearly that the cloned gene has the expected function in the biosynthesis of fungal gibberellins. Received: 28 April / 1 July 1998     

Breast carcinomas with immunocytochemical detection of aromatase in fine-needle aspirates: report of three cases

Three postmenopausal women with breast carcinoma underwent the fine-needle aspiration (FNA) preoperatively, and these specimens were stained by the antiaromatase antibody. We evaluated the identification of the aromatase immunoreactivity in breast carcinoma specimens obtained from both FNA and surgery. FNA specimens showed positive intracellular immunoreactivity of aromatase in these cases. The presence for aromatase in FNA specimens was identified with that in the surgical specimens. To our knowledge, the present cases are the first to report the aromatase staining of FNA specimens. The immunoreactivity of aromatase in FNA specimen may be useful to estimate the effectiveness of new aromatase inhibitors in patients with breast carcinoma. HUM PATHOL 32:348-351.