Microarray analysis on Phase II drug metabolizing enzymes expression in pregnant rats after treatment with pregnenolone-16alpha-carbonitrile or phenobarbital

We previously reported the expression profiles of 9 cytochrome P450 isozymes (CYPs) proteins and those of 40 CYPs genes in pregnant rat's liver, placenta and fetal liver after treatment with pregnenolone-16alpha-carbonitrile (PCN) or phenobarbital (PB). This study was carried out focusing on the gene expression profiles of Phase II drug metabolizing enzymes, Glutathione S-transferase isozymes (GSTs) and UDP-glycosyltransferase isozymes (UDPGTs). Fischer 344 (F344) pregnant rats were daily treated intraperitoneally with 50 mg/kg of PCN or 80 mg/kg of PB from 13 to 16 days of gestation (DG). They were sacrificed on 17 DG, and microarray analysis using Affymetrix Rat Expression Array 230 A was performed. Among 16 GSTs genes examined in this study, 7 genes were significantly induced in dam's liver and 3 genes in fetal liver, respectively, in the PCN-group, while 8 genes were significantly induced in dam's liver and 1 gene in fetal liver, respectively, in the PB-group. On the other hand, among 11 UDPGTs genes examined, 5 genes were significantly induced in dam's liver and 3 genes in fetal liver, respectively, in the PCN-group, while 5 genes were significantly induced in dam's liver and 1 gene in fetal liver, respectively, in the PB-group. There were no significant changes in the placenta of all groups. This is the first report of the gene expression profiles of Phase II drug metabolizing enzymes in pregnant rat and fetal livers and placenta after treatment with typical inducers of drug metabolizing enzymes.     

Simulation studies of data classification by artificial neural networks: Potential applications in medical imaging and decision making

Artificial neural networks are being investigated in the field of medical imaging as a means to facilitate pattern recognition and patient classification. In the work reported here, the effects of internal structure and the nature of input data on the performance of neural networks were investigated systematically using computer-simulated data. Network performance was evaluated quantitatively by means of receiver operating characteristic analysis and compared with the performance of an ideal statistical decision maker. We found that the relatively simple neural networks investigated in this study can perform at the level of an ideal decision maker. These simple networks were also found to learn accurately even when the training data are extremely unbalanced with respect to the prevalence of actually positive cases and to differentiate input data patterns by recognizing their unique characteristics.     

Rhythmical contractions in pulmonary arteries of monocrotaline-induced pulmonary hypertensive rats

Rhythmical contractions accompanied by an increase in cytosolic Ca2+ concentrations were produced in ring preparations of endothelium-denuded pulmonary arteries from monocrotaline-treated rats, but not in those from vehicle-treated rats, 2-3 h after a resting tension of 15 mN (150-180% of the initial wall length of the artery) was applied. The rhythmical contractions were abolished by nicardipine and ryanodine. Cyclopiazonic acid reduced the relaxation phase of the rhythmical contractions, finally leading to a sustained contraction. Similarly, apamin caused a sustained contraction, whereas charybdotoxin increased the amplitude of the rhythmical contractions. Glibenclamide had no apparent effects on them. Indomethacin and the prostaglandin H2/thromboxane A2 receptor antagonist SQ29548 abolished the rhythmical contractions and reduced the tension, but the thromboxane synthase inhibitor ozagrel had no effect. These results suggest that optimal stretch induces rhythmical contractions in the pulmonary arteries of monocrotaline-induced pulmonary hypertensive rats, to which both Ca2+ influx through voltage-operated Ca2+ channels and Ca2+ release from the endoplasmic reticulum seem to contribute. It is also suggested that small-conductance Ca(2+)-activated K+ channels participate in the relaxation phase of rhythmical contractions. Furthermore, prostaglandin H2 released from nonendothelial cells is likely to play a pivotal role in the induction of rhythmical contractions.     

Protodioscin isolated from fenugreek (Trigonella foenumgraecum L.) induces cell death and morphological change indicative of apoptosis in leukemic cell line H-60, but not in gastric cancer cell line KATO III

Protodioscin (PD) was purified from fenugreek (Trigonella foenumgraecum L.) and identified by Mass, and 1H- and 13C-NMR. The effects of PD on cell viability in human leukemia HL-60 and human stomach cancer KATO III cells were investigated. PD displayed strong growth inhibitory effect against HL-60 cells, but weak growth inhibitory effect on KATO III cells. Morphological change showing apoptotic bodies was observed in the HL-60 cells treated with PD, but not in KATO III cells treated with PD. Flow cytometric analysis showed that the hypodiploid nuclei of HL-60 cells were increased to 75.2, 96.3, and 100% after a 3-day treatment with 2.5, 5, and 10 microM PD, respectively. The fragmentation by PD of DNA to oligonucleosomal-sized fragments, that is a characteristic of apoptosis, was observed to be both concentration- and time-dependent in the HL-60 cells. These findings suggest that growth inhibition by PD of HL-60 cells results from the induction of apoptosis by this compound in HL-60 cells.     

Murine thymic nurse cell clone supports the growth of fetal thymocytes in the presence of interleukin 2.

To investigate the role of thymic nurse cells (TNC) in activation and differentiation of fetal CD4-CD8- (double-negative) thymocytes, we have co-cultured murine fetal thymocytes (14-15 days of gestation) with an established murine TNC clone. We show here that TNC induced the growth of the fetal double-negative thymocytes in the presence of recombinant interleukin 2 (rIL2). Activated fetal thymocytes markedly formed lymphocyte-TNC complexes and proliferated extensively after 5 days in the co-culture. The activated fetal thymocytes in this co-culture condition remained double negative after 10 days in culture. None of them gave rise to phenotypically and functionally competent lymphocytes during this period. TNC alone and the supernatant of TNC had no effect on activation. The presence of both TNC and rIL2 was necessary for the growth of fetal thymocytes in our system. The proliferation of fetal thymocytes was inhibited by a monoclonal antibody against mouse IL2 receptors (IL2R). The fetal thymocytes could be maintained further in this co-culture condition. The prolonged cultivation of fetal thymocytes resulted in the establishment of the fetal thymocyte line and its several clones. CD4 single-positive cells of activated fetal thymocytes first appeared 14 days after the onset of culture and their number increased, whereas CD8+ cells or CD4CD8 double-positive cells were not observed. These results indicate that fetal CD4-CD8- thymocytes underwent phenotypic change after long periods of culture. All established clones of fetal thymocytes are CD4 single positive showing lymphocyte-TNC interactions but do not express CD3 complex. Northern blot analysis detected mRNA for the gamma T cell receptor, but no messages for the delta, alpha or beta T cell receptor. Chemical cross-linking of 125I-labelled IL2 revealed that the 90-kDa band (presumably considered to be the IL2R beta chain) was clearly present in IL2-responsive fetal clones, whereas freshly isolated day 14-15 fetal thymocytes lacked the band. Taken together, TNC might be involved in the differentiation and/or expansion of murine fetal thymocytes by inducing IL2R beta chain, which forms the functional IL2R together with IL2R alpha chain and CD4, one of the T cell accessory molecules, on the cell surface through direct cell-cell interaction.     

A stereomicroscopic study of the mastopathic human breast

Summary The stereomicroscopic study of cystic disease of the breast revealed the presence of abnormal ductules near the terminal sites of the original lactiferous ducts. This alteration was the key feature of cystic disease since the involution of abnormal ductules possibly may have induced the stricture of the original duct by increasing fibrosis around the site of duct evolution. The phenomenon could contribute to the creation of cystic dilatation of distal ducts and acini. Multiple large cysts characterizing the morphology of cystic disease may represent the end stage of distended ducts and acini. Prolonged and repeated unbalanced stimulation of hormones could produce progressive and regressive alternations in the same breast. This modifies the original alteration into more pleomorphic and complicated types.

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Stereomikroskopische Studie der durch Mastopathie veränderten menschlichen MammaII. Die peripherische Form der Milchgangsevolution und ihre Beziehung zur Cystenmamma

Zusammenfassung Die stereomikroskopische Untersuchung sog. Cystenmmamen zeigt, daß die Cysten durch Ektasie der sog. Kanälchen-Endäste entstehen. Dagegen werden die Stamm-Milchgänge einer Involution zugeführt. An der Cystenbildung beteiligen sich auch die Acini. Als morphogenetische Reize kommen hormonelle Dysregulatonen in Frage. Durch die unterschiedliche Ausbildung der Kanälchen proliferation einerseits, die Involution sowie Cystenbildung andererseits resultiert ein buntes, wenig einheitliches Bild.

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Clinical Investigator, Veterans Administration Hospital, San Francisco, California, U.S.A., Former Postgraduate Student, Faculty of Medicine, Tokyo Medical and Dental University, Japan.

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This investigation was carried out at the Department of Pathology, Faculty of Medicine, Tokyo Medical and Dental University from 1958 to 1960 and was partially supported by a special research grant of the Japanese Ministry of Education.     

Effect of ambient temperature on learning

After a three week exposure at 34, 25 or 10°C, three strains of mice (C57BL/6, BALB/c and A/J) underwent a conventional active two-way avoidance task. As compared to 25°C of ambient temperature, acquisition on the first day was severely impaired in the three strains at 34°C and in the C57BL/6, and A/J mice at 10°C. Learning scores on the second day were diminished in the C57BL/6 strain at 34°C and 10°C and in the BALB/c strain at 34°C. The mechanisms of such an impairment in learning at high or low temperature are discussed.     

Rapid development of renal lesions in diabetic DBA mice infected with the D-variant of encephalomyocarditis virus (EMC-D).

Marked hyperglycaemia and renal lesions developed rapidly in DBA mice infected with 10 plaque-forming units of the D-variant of encephalomyocarditis virus (EMC-D). Renal alterations were demonstrated in the glomeruli, tubular epithelium and small vessels 2 months after infection. Glomerular changes were characterized by mesangial thickening due to an increase of basement membrane-like material in the mesangial matrix. Nodular glomerular lesions were commonly observed 3 months after infection, whereas distinct thickening of the glomerular basement membrane was rarely seen. Besides these glomerular changes, glycogen inclusions in the distal tubular epithelium and medial degeneration in the arterioles were also noticed. The EMC-D-infected DBA mouse appears to be a useful experimental model for the study of human diabetic nephropathy.     

Encephalomyocarditis (EMC) virus-induced orchitis in Syrian hamsters.

Testes of 8-week-old male Syrian hamsters which were inoculated intraperitoneally with 10(5) plaque-forming units of the D variant of encephalomyocarditis virus (EMC-D) were examined virologically and histologically. Viral replication was detected from 1 day post inoculation (1 DPI), became more prominent 3 DPI, and was no longer demonstrated 7 DPI. The weight of testis decreased in course of time and it was about 2% of that of control 6 weeks post inoculation (6 WPI). Histopathologically, degeneration and/or necrosis of germinal cells and spermatogonia were observed in many seminiferous tubules of all hamsters 3 DPI. At 7 DPI, luminal obstruction by cellular debris and subsequent replacement of them by mesenchymal cells were common in mildly atrophic tubules surrounded with inflammatory cells. Thereafter, atrophy of seminiferous tubules became severer with the lapse of time and, in addition to plasma cell infiltration, apparent increase in the number of Leydig cells was found in the interstices. No regenerative signs of germinal epithelia were detected by 6 WPI. This is the first report of EMC virus-induced orchitis.