The effect of IL-17 on the production of proinflammatory cytokines and matrix metalloproteinase-1 by human periodontal ligament fibroblasts.

OBJECTIVES: To investigate the effects of IL-17 on IL-6, IL-1ß, and matrix metalloproteinase (MMP-1) production, and to compare the MMP-1 production between the individual and combined effects of IL-1ß and IL-6 in human periodontal ligament fibroblasts (HPDLF). MATERIALS AND METHODS: Human periodontal ligament fibroblasts were cultured with IL-17 for 0.5, 1, 4, 24, 48, and 72 h, and were cultured with IL-1ß, IL-6/sIL-6R, or a combination of IL-1ß and IL-6/sIL-6R for 24 h. To measure the mRNA levels of IL-6, IL-1ß, and MMP-1, total RNA was extracted from the cultured HPDLF, and a real-time PCR analysis was performed. The protein levels of IL-6, IL-1ß, and MMP-1 in supernatants were measured using enzyme-linked immunosorbent assays (ELISAs). RESULTS: IL-17 significantly increased the expression of IL-6 and MMP-1 mRNA and protein, while IL-17 transiently increased the expression of IL-1ß mRNA. The combination of IL-1ß and IL-6/sIL-6R induced significantly higher levels of MMP-1 protein than IL-1ß alone. CONCLUSIONS: IL-17 upregulated the production of IL-6 and MMP-1 sequentially in HPDLF. IL-6/sIL-6R may enhance the effects of IL-1ß on MMP-1 production. The present results suggest that IL-17 induces MMP-1 production not only directly, but also indirectly by promoting IL-6 production, thus resulting in the degradation of collagens in the PDL.

Novel function of guanidine hydrochloride in reverse micellar extraction of lysozyme from chicken egg white.

The efficiency of guanidine hydrochloride (GuHCl) addition in the suppression of gel formation and the extraction of lysozyme during reverse micellar extraction from chicken egg white was investigated. A low concentration of GuHCl in the feed permitted the successful extraction of lysozyme in its native form without gel formation, which is perceived as a novel function of GuHCl. The highest recovery and specific activity of lysozyme were obtained at a GuHCl concentration of 0.06 M in 25 mM AOT reverse micellar extraction from 20-fold-diluted natural chicken egg white. Lysozyme and ovalbumin CD spectra in the corresponding GuHCl aqueous solutions revealed no changes in the higher order structures of the proteins. Furthermore, the specific activity of lysozyme in the feed was well preserved in the GuHCl system.

Prenatally diagnosed female prune belly syndrome associated with tetralogy of Fallot.

Female prune belly syndrome and tetralogy of Fallot were diagnosed in obstetric ultrasound screening at 13 and 28 weeks of gestation, respectively. The baby was born at 38 weeks showing abdominal distension, hypoplasia of abdominal muscles, vaginal atresia, imperforate anus and clubfeet. Radiologic and ultrasound examinations confirmed our prenatal diagnosis and revealed other abnormalities of patent ductus arteriosus, major aortopulmonary collateral arteries, bilateral vesicoureteral reflux, rectourethral fistula and hypoplasia of the right kidney. Urination has been smooth, but cystitis was observed repeatedly once a month. The infant was discharged in good condition and has been followed in the outpatient clinic.

Sister chromatid exchanges in mouse after exposure to pulse-wave ultrasound in utero.

The induction of sister chromatid exchanges (SCEs) was investigated in mice after a ten min exposure, in vivo, to 2 MHz focused, pulse-wave ultrasound with a pulse repetition rate of 1000 Hz, pulse duration of 10 microseconds. The bone marrow cells of the pregnant female mice and the fetal liver cells were analyzed. The cell cycle specific metaphase patterns were additionally evaluated. In the bone marrow cells, the mean frequencies of SCEs were 2.77 in control, 3.56 in the cells exposed to ultrasound at 586.2 mW/cm2 (spatial average temporal average, SATA); in the fetal liver cells, 2.64 in control, 3.84 in the cells exposed. The frequencies of SCEs significantly were increased by the treatment. Faster cell kinetics was observed in fetal liver cells than bone marrow cells of pregnant female. But there was no cell-growth inhibitory effect of ultrasound on both bone marrow and fetal liver cells. In fetal liver cells, the critical acoustic power was 160.0-278.9 mW/cm2 (SATA).

Change in electrocardiogram T-wave amplitude during umbilical cord compression is predictive of fetal condition in sheep.

OBJECTIVE: The purpose of this study is to assess the usefulness of the dynamic change in T/QRS ratio in fetal electrocardiograms in predicting the fetal condition when repetitive variable decelerations are seen in intrapartum cardiotocograms. STUDY DESIGN: We investigated the relationship, using linear regression and Wilcoxon's test, between T/QRS and blood gas values, catecholamine concentrations, and blood pressure during repetitive cord compression in five chronically instrumented lamb fetuses. RESULTS: T/QRS during cord compression correlated significantly (p less than 0.01) with fetal arterial pH (r = -0.7711), norepinephrine concentration (r = 0.7551), and duration of elevated blood pressure during compression (r = -0.8619). Fetal arterial pH and base excess were lower, the duration of elevated blood pressure during compression was shorter, and carbon dioxide partial pressure and catecholamine concentrations were higher in the stage with higher (greater than 0.50) T/QRS during compression (p less than 0.005). CONCLUSION: We can estimate the severity of fetal distress by measuring T/QRS near the bottom of the decelerations.

Femoral artery blood flow monitoring has distinct advantages for examining redistribution of blood flow in fetal acidosis.

Changes of blood flow in umbilical artery, carotid artery and femoral artery were examined during the progression of acidemia in fetal sheep by means of indwelling transit-time ultrasonic blood flow meters. Moreover, catecholamines in fetal blood were measured and its interrelation to the alteration in blood flow was examined. Gradually progressing fetal acidemia was induced by repeated cord compression. Umbilical blood flow showed a initial increase thereafter maintaining a plateau through the experiment, which seemed to be dependent on fetal arterial pressure. Carotid artery flow gradually increased until the arterial pH in fetal blood declined to 7.20 and remained at this level even though the acidemia further progressed. Femoral artery flow markedly decreased around fetal arterial blood pH 7.20 and its change correlated well with the plasma level of catecholamines. This change of femoral artery flow may be evaluated by examination of the flow index as well as flow volume. Redistribution of blood flow in the progression of fetal acidemia may be initiated at around fetal arterial pH 7.20 and can be detected by studying femoral artery flow.

The use of IAA to overcome interspecific hybrid inviability in reciprocal crosses between Nicotiana tabacum L. and N. repanda Willd.

Flowering hybrid plants were obtained from reciprocal crosses between N. tabacum L. (2n=48) and N. repanda Willd. (2n = 48), in which cross incompatibility and hybrid inviability were manifested. Stylar pollination and ovule culture were used to overcome the cross incompatibility of stylar barriers and ovular death. It was shown that application of 2 mg/l indole-3-acetic acid (IAA) during the growth period, from the fiveto six-leaf stage to the flowering stage, is a useful and easy method to overcome hybrid inviability.

Hematologic evaluation of growth retarded fetus by cordocentesis.

Fetal blood sampling by ultrasound-guided cordocentesis was performed in 22 IUGR fetuses in order to evaluate the fetal condition. All patients were referred to our hospital for further examination and management. In addition to hematological assessment we measured the velocity profiles of the umbilical artery with pulse-gated Doppler equipment. On the whole, pH and pO2 in the umbilical vein of IUGR fetuses were significantly lower and pCO2 higher, respectively, the corresponding values in controls. In particular, IUGR fetuses which had an etiology of maternal pre-eclampsia had fallen into hypoxemia. However, other laboratory values in IUGR did not differ from those of the controls except for the platelet count, which was low in the IUGR fetuses. Five fetuses with chromosomal abnormalities were found by analyzing the fetal blood. Since there are different etiologies inducing IUGR, fetus and umbilical artery velocimetry is not always predictive of the state of the IUGR fetus, hematological assessment is considered indispensable for further management.

Effects of a medicinal herbal liqueur, "yomeishu", on post-operative gynecological patients.

We administered 20 ml of Yomeishu (YMS) twice a day before meals for 12 weeks to 50 post-operative patients in gynecology and then inquired into their subjective 20 symptoms (sense of fatigue, insomnia, headache and heavy headedness, appetite, stomach-ache, abdominal inflation, vertigo, lumbago, etc.) The YMS group showed a significant improvement on 14 items compared with the control group. On the whole, a great improvement was observed in the YMS group with serious subjective symptoms as well, and there were significant differences for general condition, sense of fatigue, and coldness in extremities.

Biochemical evaluation of fetus with hypoxia caused by severe preeclampsia using cordocentesis.

Biochemical evaluation of the fetus using cordocentesis was performed in sixteen pregnant women with severe preeclampsia. In addition, FHR monitoring and Doppler flow velocimetry of the umbilical artery were examined in these cases before the cordocentesis. Other than blood gas and routine hematologic and biochemical examinations, catecholamine and hypoxanthine concentrations were also evaluated in some cases. According to the results it is obvious that the fetus of mother with severe preeclampsia is exposed to hypoxemia with the delta pO2, the difference from the mean in each gestation, of -22 mmHg. Especially, the prognosis of the severely growth-retarded babies with pO2 below 10 mmHg, or delta pO2 30 mmHg lower than the mean standard value, is poor. Among 16 babies, 4 with severe hypoxia or acidosis and growth retardation died perinatally. In contrast, severely growth-retarded babies without hypoxia or acidosis were alive. Since neither FHR monitoring nor Doppler velocimetry always reflect biochemical values of the fetus of a severe preeclamptic mother, hence the precise evaluation of fetal condition using cordocentesis is indispensable to determine the timing of the delivery and to prevent the neurological sequelae caused by hypoxia, especially when the fetal growth is retarded.

[Influence of pregnancy and steroid hormones on interferon production of lymphocytes].

A study was done to investigate the involvement of interferon (IFN) in immunological acceptance of fetal allograft. IFN was induced by stimulating OK-432 from lymphocyte of pregnant and nonpregnant women as well as men. There was no difference in the IFN titer in these groups. In pregnant women there was no difference between trimesters, either. In addition, we examined the influence of sex steroid hormones such as progesterone and estrogen on IFN production by the lymphocytes of women and men. Generally, each steroid hormone suppressed the production of IFN. However, the extent of suppression varied with the concentration of hormones, resulting in different suppression curves for men and women in a physiological concentration. Differences in response by men and women were also noticed when interleukin 2 was added to examine whether it could reduce the suppression by steroid hormone. This unique sensitivity of female lymphocyte to steroid hormones may play a role in successfully taking a fetal allograft in pregnancy.

[Screening for chromosomal anomalies with maternal serum alpha-fetoprotein].

Between July 1, 1986 and January 31, 1988, genetic amniocentesis was performed on 205 patients. The maternal serum alpha-fetoprotein (AFP) and amniotic fluid AFP levels were measured by enzyme immunoassay. Gestational dates were confirmed by sonography, and AFP results were expressed as multiples of the median (MOM). The median of maternal serum AFP from 15 to 17 weeks of gestation was 43.4, 62.6 and 72.5 ng/ml. Three fetuses with chromosomal anomalies were diagnosed; trisomy 21, 4p trisomy, and trisomy 18 (trisomy 18 was in one fetus of a twin pregnancy; the other fetus was normal). Maternal serum AFP levels were, 0.41, 0.49 and 1.30 MOM. Maternal serum AFP less than 0.5 MOM in normal pregnancies was 1/205 (0.5%) and less than 0.6 MOM was 9/205 (4.4%). There was no relationship between maternal serum AFP and amniotic fluid AFP levels. Our results are in agreement with the majority of the results in the literature, showing that maternal serum AFP levels are lower in association with autosomal trisomy fetuses.

Alteration of FHR pattern and cerebral metabolic rate of glucose of the fetus measured by positron emission tomography during progress of acidemia. The significance of overshoot acceleration in FHR.

In order to investigate the significance of overshoot acceleration in FHR with acidosis induced by serial cord compression, cerebral glucose metabolic rate (CMRglc) was measured in four chronically instrumented fetuses before cord compression and immediately after appearance of the overshoot acceleration in FHR. The investigation of CMRglc was carried out using positron emission tomography to assess accumulation of cyclotron-produced F18-FDG in the fetal brain. CMRglc in the fetal brain in the nonacidemic condition was 1.73 mg/100 g/min. In acidemia, with fetal arterial pH near 7.15 in which FHR showed the characteristic pattern with overshoot acceleration, CMRglc was significantly decreased to 0.73 mg/100 g/min. We concluded that, to avoid neuronal damage, it is necessary to intervene when the fetal condition deteriorates to the extent that overshoot acceleration in FHR occurs.

Three cases of fetal ascites with successful outcome and seven year evaluation of NIHF: 1981-1987.

Three pregnant women with fetal ascites were admitted to our hospital at 30, 32 and 33 weeks of gestation on diagnosis by sonogram. On sonogram, all the fetuses were initially with marked ascites but without any pleural or pericardial effusion or subcutaneous edema. Under observation by sonogram and cardiotocogram, obstetrical intervention was performed by cesarean section in all 3 cases; at 31 weeks of gestation due to notably increased ascites, at 32 weeks due to fetal distress and at 35 weeks due to development of subcutaneous edema, respectively, and the subsequent courses were uneventful. On the other hand, in our precious experience with 21 cases of NIHF over a period of 7 years, only 2 babies with NIHF have been born alive. We think fetal ascites should be considered an initial sign of NIHF and the successful outcome of these three cases were due to early diagnosis by frequent sonographic examination.

The slope of fetal heart rate deceleration is predictive of fetal condition during repeated umbilical cord compression in sheep.

The relationship between components of fetal heart rate deceleration and fetal arterial blood gas values or plasma catecholamine concentrations was investigated by repeated complete umbilical cord compression in chronically instrumented fetal lamb. Fetal arterial pH and bicarbonate levels decreased, while plasma norepinephrine and epinephrine concentrations increased more than tenfold. The slope of the descending limb of the fetal heart rate deceleration curve decreased and correlated strongly with fetal arterial pH, bicarbonate, and logarithmic plasma norepinephrine and epinephrine concentrations. Fetal arterial pH and bicarbonate levels were significantly lower in the group with lower fetal heart rate deceleration slope, and a greater plasma catecholamine concentration in this group suggested a redistribution of blood flow to vital organs. Therefore, during repeated umbilical cord compression, the fetal acid-base and hormonal state was predicted by the fetal heart rate deceleration slope. This relationship may be applicable to human fetuses in the diagnosis of fetal distress caused by umbilical cord compression during labor.

Studies on the chromosome by scanning electron microscope--a new trial with long-term Giemsa staining.

Chromosomes, which can be seen under a light microscope (LM) as bands, are too thin to see with a scanning electron microscope (SEM). We noticed that chromosomes increase in thickness with Giemsa staining, so that scanning microscopic specimens could be prepared by long-time Giemsa staining for SEM observation, and in the present study the following results were obtained. 1) When the duration of the trypsin treatment was kept constant, and the Giemsa staining was performed from 30-min to 24-h, it was found that the thickness of the chromosomes increased with the length of treatment time, and the fibrous structures were clearly seen. 2) While a sufficient thickness of the chromosomes was obtained with the long-time Giemsa staining, the bands became unclear as the time of staining was extended. 3) A long-time Giemsa staining without trypsin treatment showed more "highly packed" fibers, so that the trypsin treatment was felt to be related to the dispersion of the fibers.

Alteration of FHR pattern associated with progressively advanced fetal acidemia caused by cord compression.

In order to investigate changes in FHR when a fetus suffers acidemia, we produced progressively advanced acidemia in lamb fetus by intermittently repeated cord compression. FHR was monitored throughout the study. FHR patterns were classified into five characteristic types as fetal arterial pH fell from around 7.35 to below 6.90. It was confirmed by studies involving catecholamine release and the administration of drugs such as atropine sulfate, alpha or beta adrenergic antagonists that sympathetic and parasympathetic regulation was deeply involved in the changes. Among the patterns, type 4, in which decreased amplitude of the initial drop and hypoxia-induced deceleration was followed by overshoot acceleration, took place during acidemia at arterial pH below 7.15. In conclusion, the appearance of type 4 FHR indicates a deteriorating state in human fetus as well, induced by repeated cord compression, and obliges us to deliver the fetus as soon as possible.