Infrequent genetic alterations of the PTEN gene in Japanese patients with sporadic prostate cancer

Prostate cancer is a major cause of cancer death among elderly men in America, Europe, and Japan. However, the molecular mechanism of carcinogenesis is not yet well characterized. Frequent loss of heterozygosity (LOH) on chromosome 10q was reported in prostate cancer, and a candidate tumor suppressor gene, PTEN, was isolated on chromosome band 10q23.3. To investigate the genetic alterations of PTEN, we examined 45 primary prostate cancer specimens. LOH at the PTEN locus was observed in two (11.1%) of 18 tumors. However, no mutations were observed in any of the primary prostate cancers. These data suggest that mutation of the PTEN gene does not play a major role in prostate carcinogenesis of Japanese patients. Received: February 6, 1998 / Accepted: July, 3, 1998     

Orientational behavior of side-chain liquid-crystalline polysiloxanes deduced from measurements of second harmonic generation

Side-chain functionalized polysiloxanes were prepared via polymer-analogous esterification of poly[(3-chloroformylpropyl)methylsiloxane] with 4-(4-hydroxyphenylazo)nitrobenzene ( P1 ), 4-[4-(ω-hydroxyalkyloxy)phenylazo]nitrobenzene ( P2 – P4 ), 4-{4-[N-(2-hydroxyethyl)-N-methyl]anilinoazo}nitrobenzene ( P5 ), 4-(4-hydroxypiperidino)nitrobenzene ( P6 ), or 4-[4-(2-hydroxyethyl)piperidino]nitrobenzene ( P7 )., P1 , P3 , P4 and P5 exhibit liquid crystallinity, as deduced from differential scanning calorimetry, polarized microscopic observations and X-ray diffraction measurements. The liquid-crystalline phase of P1 and P5 is a nematic phase, and that of P3 and P4 is a smectic one. The second harmonic generation (SHG) measurement of a spin-coated film of P1 was carried out by the Maker fringe method using a Q-switched Nd:YAG laser (1064 nm). The SHG profile after the heat treatment of a spin-coated film suggests a perpendicular orientation of the mesogenic molecules to the glass substrate. The SH light intensity of a corona-poled film was 20-fold higher than that of a film which was only heated, though no differences were observed in their UV-vis absorption spectra. These findings suggest that the mesogenic-molecular dipole moments are aligned to the same direction in the crystalline or liquid-crystalline phase by a poling treatment.     

Cloning and characterization of the cDNA encoding the HA protein of a hemagglutination-defective measles virus strain

cDNA clones corresponding to the mRNA for the hemagglutinin of the hemagglutination-defective strain AK-1 of measles virus were isolated and characterized. Compared with the prototype Edmonstron strain, 60 nucleotide substitutions that resulted in 18 amino acid changes were detected. An additional potential N-linked glycosylation site was added by point mutation, which was supported by the observation that the hemagglutinin of the AK-1 strain was stained more heavily after NaDodSO₄PAGE and periodic acid-Schiff (PAS) staining than the Edmonston strain. Computer-assisted analysis revealed that three reverse turns in the secondary structure had disappeared in the hemagglutinin of the AK-1 strain. Moreover, one of these structural changes occurred in the closely glycosylated region at amino acid residues 168–240, which appeared to be a biologically important functional domain. The isoelectric point calculated from the predicted amino acid sequence became about 1 pH unit more basic in the AK-1 strain than the Edmonston strain. This present study is the first sequence analysis of the hemagglutinin gene in a hemagglutination-defective strain of the measles virus.     

Early degeneration of the epithelial cells in the initial segment of the epididymal duct in mice after efferent duct cutting

Mouse epididymides were examined by light and electron microscopy 6, 12, 24 h, 1.5, 2, 3, and 5 days after efferent duct cutting when the mice were 60 days of age. Six hours after operation, the principal cells in the initial segment of the epididymal duct began to degenerate following the disappearance of intraluminal spermatozoa. The degenerated cells increased rapidly and reached their greatest number 24 h, and then decreased to smaller numbers until 48 h. These degenerative changes were followed by the appearance of macrophages in the epithelium, which were first seen 12 h after operation, becoming very frequent at one and 1.5 days, and then decreasing to become rare at 3 and 5 days. The macrophages phagocytosed the degenerated principal cells. The degenerated principal cells were also ingested and digested by undegenerated principal cells and basal cells. The intraepithelial mitotic figures had almost disappeared at 24 h but were frequently observed at 2 days and more so at 3 days. They returned to normal numbers at 5 days. The volume of the initial segment was decreased to one third until the third day. The principal cells in this segment became similar in both light-microscopic appearance and ultrastructures to the cells in the next segment 5 days after efferent duct cutting. The changes were localized only in the initial segment.     

Enhancement of lymphocyte response to PHA by lysosomal enzymes from polymorphonuclear leukocytes of RA joint fluid

The effect of polymorphonuclear leukocyte (PMN) granule lysates obtained from joint fluid of RA an the in vitro DNA synthesis of PHA-stimulated autologous lymphocytes from joint fluid was studied. Lymphocytes were cultured for 3 days with or without PMN lysates in 2 ml of RPMI-1640 supplemented with 10% heat-inactivated fetal calf serum (FCS). The lymphocytes were stimulated with phytohemagglutinin (PHA-M). The DNA synthesis was measured by counting the [³H]thymidine incorporation. Lymphocytes from RA joint fluid stimulated with PHA-M showed 19,466±987 cpm (mean±SE per 10⁶ cells in the absence of PMN lysates. Upon addition PMN lysates to the PHA-stimulated lymphocytes, the maximum in vitro DNA synthesis increased to 44,877±1338 cpm. The enhancing effect of PMN lysates was abolished by plasma inhibitors or by passage through a column of protease inhibitor (Trasylol). It was concluded, therefore, that the enhancing effect of PMN lysates on PHA-stimulated lymphocytes may be associated with lysosomal proteases. Based on experiments using separated T and B lymphocytes, the enhancing effect of PMN lysates was considered to result from the activation of T lymphocytes. The results obtained in the present study suggest an important role for lysosomal proteases in the perpetuation of rheumatoid synovitis.     

Significant role of interleukin-8 in pathogenesis of pulmonary disease due to Mycoplasma pneumoniae infection

We found elevated levels of interleukin-8 in pleural fluid samples from patients with pleural effusion and with a sustained fibrotic change of the lung due to Mycoplasma pneumoniae infection. This result suggests a critical role of interleukin-8 in the pathogenesis of a certain type of pulmonary disease caused by M. pneumoniae.     

Correlation dimension analysis of the artificial circulation

Artificial circulation has been analyzed by decomposing it into parts. However, the sum of the decomposed parts is not equal to the whole system, especially in nonlinear dynamic systems such as biological systems. To evaluate prosthetic circulation as an entity, not as decomposed parts, nonlinear mathematical analytic techniques, including fractal dimension analyzing theory, were used. Two pneumatically actuated ventricular assist devices were implanted as biventricular bypasses (BVB) in chronic animal experiments using four healthy adult goats. For comparison between natural and prosthetic circulation in the same experimental animals, the BVB-type complete prosthetic circulation model with ventricular fibrillation was adopted. All hemodynamic parameters with natural and prosthetic circulation were recorded under awake conditions and calculated by a personal computer system. By the use of nonlinear mathematical techniques, time-series data of the hemodynamics were embedded into the phase space, and correlation dimension analysis was performed to evaluate the reconstructed attractor. Our results suggest that the correlation dimension of the arterial blood pressure does not linearly increase according to the increase of the embedding dimension, even during artificial circulation, suggesting those are the fractal time series data. Dimensional analysis of the hemodynamics revealed that lower dimensional fractal dynamics were observed during prosthetic circulation. Fractal time series data are suggested to have robustness and error resistance. Thus, our results suggest that the circulatory regulatory system with the artificial heart may have these desirable characteristics. Accepted: July 14, 1995     

Isolation, characterization, and disruption of dnr1, the areA/nit-2-like nitrogen regulatory gene of the zoophilic dermatophyte, Microsporum canis.

A homolog of the major nitrogen regulatory genes areA from Aspergillus nidulans and nit-2 from Neurospora crassa was isolated from the zoophilic dermatophyte, Microsporum canis. This gene, dnr1, encodes a polypeptide of 761 amino acid residues containing a single zinc-finger DNA-binding domain, which is almost identical in amino acid sequence to the zinc-finger domains of AREA and NIT-2. The functional equivalence of dnr1 to areA was demonstrated by complementation of an areA loss-of-function mutant of A. nidulans with dnr1 cDNA. To further characterize this gene, dnr1 was disrupted by gene replacement based on homologous recombination. Of 100 transformants analyzed, two showed the results expected for replacement of dnr1. The growth properties of the two dnr1(-) mutant strains on various nitrogen sources were examined. Unlike the A. nidulansareA(-) mutant, these dnr1(-) mutants showed significantly reduced growth on ammonia, a preferred nitrogen source for fungi. These mutant strains were also able to utilize various amino acids for growth. In comparison with wild-type M. canis, the two dnr1(-) mutants showed reduced growth on medium containing keratin as the sole nitrogen source. This is the first report describing successful production of targeted gene-disrupted mutants by homologous recombination and their phenotypic analysis in dermatophytes.