Ambulatory BP monitoring and clinic BP in predicting small-for-gestational-age infants during pregnancy.

The significance of ambulatory blood pressure (ABP) monitoring during pregnancy has not been established. We performed a prospective study to elucidate whether ABP measures are associated with small-for-gestational-age birth weight (SGA). We studied 146 pregnant women who were seen for maternal medical checkups or suspected hypertension. ABP monitoring was performed for further assessment of hypertension. The outcome measure was SGA. The subjects were classified by their medical history and ABP as having preeclampsia or gestational hypertension (n=68 cases), chronic hypertension (n=48) or white-coat hypertension (n=30). There were 50 (34.2%) cases of SGA by the fetal growth reference standard. In multivariable logistic regression analyses adjusting for age, body mass index, the presence of prior pregnancy, current smoking habit and the use of antihypertensive medications, 24-h SBP (per 10 mm Hg (odds ratio (OR): 1.74; 95% confidence interval (CI): 1.28-2.38; P<0.001)) was more closely associated with SGA than clinic BP (OR: 1.40; 95% CI: 0.92-2.13; P=0.11). The results were essentially the same if 24-h BP was replaced by awake or sleep SBP. Ambulatory diastolic BP showed the same tendency. However, abnormal circadian rhythm was not associated with the outcome. In conclusion, ambulatory BP monitoring measures performed during pregnancy were more closely associated with SGA than clinic BP.

PP058. The additive effect of plasma levels of sFlt-1/PLGF ratio following the risk classification using both blood pressure levels and uterine artery blood flow impedance in the second trimester on the later occurrence of preeclampsia.

INTRODUCTION: The multivariate model including circulating levels of soluble fms-like tyrosine kinase 1 (sFlt-1)/placental growth factor (PlGF) ratios, maternal factors, blood pressure (BP) levels and uterine artery (UtA) doppler in the first to second trimester has been reported to be clinically useful to predict PE more accurately. However, the effects of levels of sFlt-1/PlGF ratio after the stratification of women using two major risk factors for PE, BP levels and UtA blood flow imdedance (BFI) have not been evaluated. OBJECTIVES: Our aim was to evaluate the additive effect of plasma levels of sFlt-1/PlGF ratio following the risk classification using both mean blood pressure (MBP) levels and the combination of two UtABFI, mean pulsatility index (mPI) and mean notch depth index (mNDI), on the later occurrence of PE. METHODS: 1161 women were recruited into a prospective cohort study during 2004 and 2008. Clinical BPs were measured twice during 16 and 23 weeks, UtA doppler was performed twice during 16 and 23 weeks, and the mPI and mNDI was measured. Plasma samples were drawn once at 20-23weeks, and were stored at -20°C until use. The levels of sFlt-1/PlGF ratio were measured by automated electrochemiluminescent immunoassay (Roche Diagnostics K.K.). The cutoff value of mean BP (MBP) was determined as 91.3mmHg using ROC curve, and that of sFlt-1/PlGF ratio was 13.0, the 97.5th percentile of log10(sFlt-1/PlGF) at 20-23 weeks in normal pregnant women. If the mPI was <90th percentile of the gestational-age specific reference range of mPI, or the mNDI was <90th percentile of the gestational-age specific reference range of mNDI, we defined that the UtABFI was low; if the mPI was ⩾90th percentile and the mNDI was ⩾90th percentile, we defined that the UtABFI was high. RESULTS: When women were stratified to 4 groups: low BP and low UtABFI, high BP but low UtABFI, high UtABFI but low BP, high BP and high UtABFI, the PPVs were 0.7%, 6.9%, 6.2% and 39.1%, respectively. In women with low BP and low UtABFI, the high sFlt-1/PlGF changed the PPV to 11.1%; the interval from the sampling to the onset of PE in women with high sFlt-1/PlGF ratio was significantly shorter than in those with low sFlt-1/PlGF ratio (mean±SD [weeks]: 5.9±1.5 vs. 16.5±2.1, p<0.01). In women with high BP but low UtABFI, the high sFlt-1/PlGF changed the PPV to 18.2%; the interval from the sampling to the onset of PE in women with high sFlt-1/PlGF ratio was significantly shorter than in those with low sFlt-1/PlGF ratio (8.0±5.7 vs. 12.6±3.7, p<0.05). Although these effects of sFlt-1/PlGF ratio on the occurrence of PE were not confirmed in women with high BP and high UtABFI, PE occurred in all women with three risk factors (5/5), and the interval from the sampling to the onset of PE was6.8±4.1 weeks. CONCLUSION: Women with both high BP and high UtABFI, especially those with additional risk of high sFlt-1/PlGF ratio, were the highest risk of PE. In the lowest risk group of low BP and low UtABFI, the addition of sFlt-1/PlGF ratio improved the PPV and the interval from the sampling to the onset of PE. These results clearly indicated the clinical importance of measuring sFlt-1/PlGF ratio in addition to BP levels and UtABFI in all pregnant women in the second trimester.

Prediction of ovarian reserve based on day-3 serum follicle stimulating hormone concentrations during the pituitary suppression cycle using a gonadotropin releasing hormone agonist in patients undergoing in vitro fertilization-embryo transfer.

Satisfactory results following in vitro fertilization-embryo transfer (IVF-ET) treatments depend on retrieving an appropriate number of mature oocytes without causing the development of ovarian hyperstimulation syndrome (OHSS). The present study was carried out to investigate whether the ovarian reserve is predictable based on the day-3 serum concentration of follicle stimulating hormone (FSH) during the pituitary suppression cycle using a gonadotropin releasing hormone (GnRH) agonist (defined as day-3 FSH) in patients undergoing IVF-ET treatment. Day-3 FSH before the administration of gonadotropin was assessed in 72 IVF-ET cycles from 59 infertile women. The mean+/-SD of day-3 FSH, the total amount of FSH plus human menopausal gonadotropin (hMG) administered, and the total number of oocytes retrieved was 5.5+/-2.6 mIU/ml, 2834.2+/-1236.5 IU and 7.7+/-5.8, respectively. There were significant correlations between day-3 FSH and the total amount of FSH-hMG administered (p < 0.001), and day-3 FSH and total number of oocytes retrieved (p < 0.001). There was a significant difference of day-3 FSH between patients who subsequently conceived (4.4+/-1.3 mIU/ml) and those who did not conceive (6.1+/-2.9 mIU/ml) (p = 0.001). There was also a significant difference of day-3 FSH between patients who developed moderate or severe OHSS (4.5+/-1.2 mIU/ml) and those who did not (5.9+/-2.8 mIU/ml) (p = 0.003). Receiver-operator characteristic curve analysis showed that the significant cut-off point for day-3 FSH for predicting ovarian reserve was 5.25 mIU/ml. These findings indicate that day 3-FSH is usefulfor predicting ovarian reserve during the pituitary suppression cycle using a GnRH agonist in patients undergoing IVF-ET.

Isolated pericardial effusion and transient abnormal myelopoiesis in a fetus with Down's syndrome.

Isolated pericardial effusion was detected in a fetus at 34 weeks of gestation. A male infant weighing 2,044 g was born by cesarean section because of a non-assuring fetal heart rate pattern at 35 weeks of gestation. Transient leukocytosis (36,100/microl) with 49% blast cells was seen in this neonate. The infant's karyotype was 47, XY + 21. The pericardial effusion disappeared after treatment with prednisolone at a dose of 2 mg/kg/day. Hypothyroidism was subsequently found. Thus, the subject patient with Down's syndrome developed isolated pericardial effusion, transient abnormal myelopoiesis (TAM), and hypothyroidism. Because more than 20% of the infants with TAM and Down's syndrome develop acute nonlymphocytic leukemia in early childhood, he is being closely observed.

Local injection of OK432 can augment the TH1-type T-cell response in tumor-draining lymph node cells and increase their immunotherapeutical potential.

The effect of local injections with streptococcal preparation OK432 on the therapeutical potential of tumor-draining lymph node (LN) cells was investigated in mice. Peritumoral injections with OK432 on days 2, 4, 6, 8 and 10 showed no effect on the in vivo growth of s.c. inoculated B16F10 melanoma. The B16F10-draining OK432-treated LN cells, however, showed a high level of anti-B16F10 cytolytic activity after an in vitro culture first with both anti-CD3 monoclonal antibody (MAb) and activated B cell blasts, and subsequently with interleukin (IL)-2 without in vitro restimulation. Such in vitro expanded LN cells showed a remarkable antitumor effect against pulmonary metastasis of B16F10 melanoma, even without the concurrent administration of IL-2. In addition, the therapeutical protocol was also found to be moderately effective against poorly immunogenic MCA fibrosarcoma, and the in vivo antitumor effect was specific to the tumor from which the LNs were harvested. Interestingly, 2 kinds of comparative analyses of the cytokines revealed that the B16F10-bearing state induced the draining LN cells to develop a Th2-type response. However, the OK432 treatment was able to effectively augment their Th1-type response. Collectively, our results suggest that peritumoral injections with OK432 significantly increased the therapeutical potential of the tumor-draining LN cells by augmenting their Th1-type response.

The augmenting effect of OK432-stimulated B cells on the in vitro generation of anti-tumor cytotoxic T lymphocytes from tumor-draining lymph node cells: the possible role of interleukin-12.

Effect of a local injection with a streptococcal preparation OK432 on the in vitro generation of anti-tumor cytotoxic T lymphocytes (CTLs) from tumor-draining lymph nodes (LN) was investigated. A peritumoral injection with OK432 on days 2, 4, 6 and 8 significantly increased both the total cell number and the proportion of B cells in the draining LN cells on day 10 after a subcutaneous inoculation with B16 melanoma. In an in vitro proliferative assay, OK432 showed a stimulatory effect on both normal splenic T and B cells. In a cytolytic assay, the OK432-injected B16-draining LN cells showed a higher level of anti-B16 CTL activity than the B16-draining LN cells after in vitro restimulation. This augmenting effect of OK432 was dependent on the B cells. Moreover, nonadherent cells from the OK432-injected B16-draining LN cells showed a low but significantly higher level of anti-B16 CTL activity than those from the B 16-draining LN cells after in vitro restimulation, whereas this augmenting effect of OK432 was abolished by the in vitro addition of anti-interleukin (IL)-12 monoclonal antibody. Collectively, these findings suggest that the augmenting effect of a local injection with OK432 on the potential of tumor-draining LN cells to turn into anti-tumor CTLs after in vitro restimulation was at least in part due to IL-12 derived from the OK432-stimulated B cells.

Effects of interleukin-12 on in vitro culture with interleukin-2 of regional lymph node lymphocytes from lung cancer patients.

In the present study, we carried out a functional analysis of regional lymph node lymphocytes (RLNL) from patients with lung cancer after in vitro activation by interleukin-2 (IL-2) and interleukin-12 (IL-12). IL-12 (100 U/ml) enhanced both the proliferation and cytotoxic activity of RLNL in a culture with low doses of IL-2 (5-10 JRU/ml). After comparing an RLNL culture with a low dose of IL-2 alone, a higher proportion of CD8+ cells and CD56+ cells and a lower proportion of CD4+ cells were found in the culture with both IL-12 and a low dose of IL-2. Such a combination of the cytokines effectively activated RLNL in terms of the expression of IL-2 receptors. In the culture condition of IL-12 and a low dose of IL-2, a synergistic effect was observed in the production of such cytokines as interferon gamma, tumor necrosis factor alpha (TNF alpha), and TNF beta, as well as in tumor cytotoxicity. However, the addition of IL-12 inhibited the cytotoxicity of RLNL in the culture with a high dose of IL-2 (100 JRU/ml). This inhibition is considered to be partially due to the endogenous production of TNF alpha by lymphocytes, because the neutralization of TNF alpa bioactivity partially restored the cytotoxic activities of RLNL. Furthermore, in the presence of hydrocortisone, IL-12 synergistically enhanced the cytotoxic activity of RLNL cultured with a high dose of IL-2. These results provide useful information about the improvement of adoptive immunotherapy against cancer using RLNL.

A voltage-dependent chloride channel from Tetrahymena ciliary membrane incorporated into planar lipid bilayers.

Membrane vesicles from cilia of Tetrahymena thermophila were incorporated into a planar phospholipid bilayer membrane, and single-channel currents across the planar membrane were recorded under voltage-clamp conditions. A novel and reproducible chloride channel was observed when a mixture of phosphatidylethanolamine and phosphatidylcholine was used to form the planar lipid membrane but not when acidic phospholipid mixtures such as asolectin or a mixture containing phosphatidylserine. Using symmetrical 100 mM KCl solutions, the single-channel conductance of the fully open state (O1) was 73.1 pS, with sub-level (O2) conductance of 9.0 pS. The permeability ratio Pc1/Pk was calculated as 3.7, according to the Goldman-Hodgkin-Katz current equation. This channel exhibited characteristic voltage-dependent burst activities. With an increase in membrane potential, the lifetimes of both the burst and interburst states decreased. In the burst state, the frequency of transition between the O1 and O2 states was also voltage-dependent, mainly due to the decrease in the lifetime of the O1 state, with an increase in membrane potential. In addition, channel activity was inhibited by indanyloxyacetic acid-94 (IAA-94), an inhibitor of epithelial chloride channels.

Specific immunotherapy with tumour-draining lymph node cells cultured with both anti-CD3 and anti-CD28 monoclonal antibodies.

For providing costimulatory signals, we utilized anti-CD28 monoclonal antibody (mAb) for the in vitro culture of tumour-draining lymph node (LN) cells. The proliferation of B16 melanoma-draining LN cells in the culture with anti-CD3 mAb was remarkably enhanced by the addition of anti-CD28 mAb. In culture with both anti-CD3 and anti-CD28 mAb, the B16-draining LN cells produced a higher level of interferon-gamma, but not interleukin-4, than with anti-CD3 mAb alone. The B16-draining LN cells efficiently expanded in the culture with both anti-CD3 and anti-CD28 mAb and subsequently with a low dose of IL-2 (anti-CD3 plus anti-CD28/IL-2). The expanded cells consisted predominantly of CD8+ T cells and showed a specific cytolytic activity, in a major histocompatibility complex (MHC) class I-restricted manner, even without in vitro restimulation. In addition, the adoptive transfer of the B16-draining LN cells, expanded in the culture protocol of anti-CD3 plus anti-CD28/IL-2, showed a significant anti-tumour effect against metastatic B16 melanoma in combination with IL-2. The cured mice thus acquired a specific protective immunity. Moreover, this protocol was also moderately effective against poorly immunogenic 3LL carcinoma. Overall, our results suggest the potential for another immunotherapeutic strategy based on 'the costimulatory theory' other than vaccination with B7-transfected tumour cells.

Conditioning-specific modification of the rabbit's unconditioned nictitating membrane response.

Robust classical conditioning modifies responding to the unconditioned stimulus (US) in the absence of the conditioned stimulus (CS), a phenomenon the researchers called conditioning-specific reflex modification. Unconditioned responses (URs) to periorbital stimulation varying in intensity and duration were assessed before and after 1, 3, or 6 days of paired, explicitly unpaired, or no presentations of tone and electrical stimulation. After 3 days of pairings, conditioned responding (CRs) reached 94%, and there was an increase in latency to the peak of URs. The peak latency increase was replicated in a second experiment where rabbits reached asymptotic conditioning during 6 days of pairings. There was also a conditioning-specific increase in the amplitude of URs. There were no UR changes as a function of low level of CRs following 1 day of pairings. Data suggest that there are learning-specific changes in pathways mediating the US/UR, as well as in those mediating the CS/CR.