Drug-induced pneumonitis caused by sulfamethoxazole, trimethoprim during treatment of Pneumocystis carinii pneumonia in a patient with refractory ulcerative colitis

We recently treated a patient with intractable ulcerative colitis complicated with Pneumocystis carinii pneumonia in whom sulfamethoxazole/trimethoprim caused pneumonitis. The pneumonitis was difficult to differentiate from worsening of the infection or the appearance of another opportunistic infection. The patient's history of sulfasalazine (sulfonamide)-induced pneumonitis made diagnosis possible. The CD4/CD8 ratio of lymphocyte subsets in bronchoalveolar lavage fluid was decreased at the diagnosis of Pneumocystis carinii pneumonia and this ratio had increased when drug-induced pneumonitis was diagnosed. Topical administration of beclomethasone dipropionate by enema was a safe and effective for the treatment of such a compromised patient with active colitis.     

Induction of ectopic corticotropic tumor in mouse embryos by exo utero cell transplantation and its effects on the fetal adrenal gland

To establish an in vivo experimental system for developmental endocrinology research, AtT-20 cells, a corticotropic tumor cell line, were transplanted by exo utero manipulation into mouse embryos on embryonic day 14. The induced tumor secreted ACTH in situ, and the circulating ACTH level was elevated. This was the first model for studying the regulation of ACTH in the mouse fetal adrenal in vivo and the first continuous ACTH treatment model in rodent fetuses. The changes in the adrenal gland from the tumor-induced embryos were analyzed by light microscopic morphometry, immunohistochemistry for steroidogenic enzymes, and electron microscopy. In the treated adrenal, the volume of the inner cortical zone was significantly larger than that in controls. In the inner zone, cell density was decreased, and average cell size was increased, whereas bromodeoxyuridine-incorporation was not increased. The enlarged inner zone cells expressed an enhanced level of cytochrome P45011beta, the corticosterone-synthesizing enzyme, and the serum corticosterone level was increased. Electron microscopy showed an active form of the organelles involved in steroidogenesis. These findings indicate that ACTH stimulates both adrenocortical hypertrophy and steroidogenesis in fetal mice. Potential perspectives of the novel paradigm in this research for molecular developmental endocrine study are discussed.     

Improvement of bitumen-waste product in leachability—Leachability of bitumen product containing BWR's evaporator concentrate

The method has been developed to improve the bitumen product which incorporates an evaporator concentrate from a BWR, with respect to the swelling and leaching. The leachability of the product has been measured by the method recommended by the IAEA. The swelling of the product is successfully prevented by the addition of calcium chloride. The specimen containing the waste up to Wa/B (Weight ratio of Na₂SO₄ + CaCl₂ and bitumen) = 60:40 shows no pronounced swelling, when it is immersed in water. The cumulative fractions of ¹³⁷Cs and ⁶⁰Co leached from a specimen which does not contain CaCl₂ are 0.65 and 0.2 at the leaching time of 30 days. On the hand, the corresponding value at 100 days for the specimen with calcium chloride addition is 5 × 10^?4 for ¹³⁷Cs and 1 × 10^?4 for ⁶⁰Co. The coating of the specimen surface with a fresh bitumen (5 mm thickness) reduces the leachability further. These results indicate that this method is effective to improve the bitumen product incorporating BWR's evaporator concentrate.     

Acidophilic xylanase from Aureobasidium pullulans: efficient expression and secretion in Pichia pastoris and mutational analysis

A yeast-like fungus Aureobasidium pullulans var. melanigenum strain ATCC 20524 produces an extracellular acidophilic endo-1,4-beta-xylanase with an optimum pH of 2.0 [Ohta et al., J. Biosci. Bioeng., 92, 262-270 (2001)]. The xynI cDNA encoding the precursor protein (XynI) was expressed in the methylotrophic yeast Pichia pastoris under the control of the alcohol oxidase I gene promoter. The 34 amino acid prepro-signal peptide of the A. pullulans XynI directed the efficient secretion of 178 mg of active xylanase per liter of the culture medium. The secretion level of the xylanase with its own signal peptide was comparable to that of the mature protein fused to the prepro leader from Saccharomyces cerevisiae alpha-mating factor and twofold higher than that of the mature protein fused to the pre-type signal peptide from P. pastoris acid phosphatase. The N-terminal amino acid sequence and the apparent M(r) of 24 kDa of the secreted recombinant protein indicated the native-like processing of the A. pullulans XynI signal sequence in P. pastoris. The three-dimensional model and mutational analysis of the xynI gene product showed that Asp-73 and Glu-157 residues located at the upper and lower edges of the active site cleft, respectively, play a significant role in its low pH optimum.     

The protein encoded by cancer/testis gene D40/AF15q14 is localized in spermatocytes, acrosomes of spermatids and ejaculated spermatozoa

We have previously identified and cloned a human gene, D40, that is preferentially expressed in testis among normal organs, while it is widely expressed in various human tumor cell lines and primary tumors derived from different organs. In this report, we have examined the expression and localization of this protein in human testis with an antibody specific to D40 protein. In Western analyses, the anti-D40 antibody recognized a major band with a molecular mass of 300 kDa and a minor band of 250 kDa. These bands were not observed in the testis lysates from patients with Sertoli-cell-only syndrome and with Kleinfelter syndrome, who lack germ cells of the testis, indicating that D40 protein is expressed in the germ cells of normal testis. Immunohistochemical studies have revealed that D40 protein is highly expressed in spermatocytes and in the pre-acrosome of round spermatids. In the acrosome, D40 protein expression is observed not inside but outside the acrosome membrane. This is consistent with the finding that the amino-acid sequence at the amino terminal of the D40 protein lacks a hydrophobic signal peptide that is required for proteins to translocate to the membrane. Expression of D40 protein is observed in the acrosome of ejaculated spermatozoa as well, although the level is low compared with that in the pre-acrosome of spermatids. These results suggest that D40 protein plays important roles in spermatogenesis, especially in the formation and maintenance of the acrosome.     

Mutants of the nematode Caenorhabditis elegans that are defective specifically in their attraction to cycloheximide

The nematode Caenorhabditis elegans exhibits chemotaxis toward a wide variety of chemicals including water-soluble molecules and volatile organic compounds. We have previously discovered that C. elegans wild-type strain N2 is strongly attracted by cycloheximide which has long been known as a bitter tastant for humans and other mammals. We describe here the isolation and initial characterization of the first mutants which were defective specifically in their attraction to cycloheximide. In our screenings, we selected two mutants that were defective in chemotaxis to cycloheximide but normal in their attraction to NH4Cl and histidine. These mutants also avoided quinine hydrochloride, CuSO4 and high concentrations of NaCl similar to the wild-type strain N2. Furthermore, no observable defect was detected in their attraction to volatile odorants such as isoamyl alcohol and diacetyl. Dye-filling experiments suggested that they have no morphological defect in the sensory endings of the amphid neurons.