bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clomiphene, an Ovulation-Inducing Agent, Mobilizes Intracellular Ca<sup>2+</sup> and Causes Extracellular Ca<sup>2+</sup> Influx in Bladder Female Transitional Carcinoma Cells
Authors: Jan Chung-Ren , Yu Chia-Cheng , Huang Jong-Khing 
Published: April 2003

<i>Background/Methods:</i> The effect of clomiphene, an ovulation-inducing agent, on cytosolic free Ca<sup>2+</sup> levels ([Ca<sup>2+</sup>]<sub>i</sub>) in populations of BFTC human bladder cancer cells was explored by using fura-2 as a Ca<sup>2+</sup> indicator. <i>Results:</i> Clomiphene at concentrations between 10 and 75 µ<i>M</i> increased [Ca<sup>2+</sup>]<sub>i</sub> in a concentration-dependent manner and the signal saturated at 50 µ<i>M</i>. The [Ca<sup>2+</sup>]<sub>i</sub> signal was biphasic with an initial rise and a slow decay. Ca<sup>2+</sup> removal inhibited the Ca<sup>2+</sup> signal by about 40–50% in maximum [Ca<sup>2+</sup>]<sub>i</sub>. Adding 3 m<i>M</i> Ca<sup>2+</sup> increased [Ca<sup>2+</sup>]<sub>i</sub> in cells pretreated with 50 µ<i>M</i> clomiphene in Ca<sup>2+</sup>-free medium, suggesting that clomiphene induced capacitative Ca<sup>2+</sup> entry. In Ca<sup>2+</sup>-free medium, pretreatment with 50 µ<i>M</i> brefeldin A (to disrupt the Golgi complex Ca<sup>2+</sup> store), 1 µ<i>M</i> thapsigargin (to inhibit the endoplasmic reticulum Ca<sup>2+</sup> pump), and CCCP (to uncouple mitochondria) inhibited 85% of clomiphene-induced intracellular Ca<sup>2+</sup> release. Conversely, pretreatment with 50 µ<i>M</i> clomiphene in Ca<sup>2+</sup>-free medium abolished the [Ca<sup>2+</sup>]<sub>i</sub> increase induced by brefeldin, thapsigargin or CCCP. The intracellular Ca<sup>2+</sup> release was unaltered by inhibiting formation of inositol-1,4,5-trisphosphate (IP<sub>3</sub>) with 2 m<i>M</i> 1-(6-((17β-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122; a phospholipase C inhibitor). <i>Conclusion:</i> The [Ca<sup>2+</sup>]<sub>i</sub> increase induced by 50 µ<i>M</i> clomiphene was not affected by 10 µ<i>M</i> of nifedipine, verapamil or diltiazem. Collectively, the results suggest that clomiphene releases intracellular Ca<sup>2+</sup> in an IP<sub>3</sub>-independent manner and also activates extracellular Ca<sup>2+</sup> influx.
Insulin Secretion and Acetylcholinesterase Activity in Monosodium <i>L</i>-Glutamate-Induced Obese Mice
Authors: Lucinei Balbo Sandra , Gravena Clarice , Bonfleur Maria Lucia , de Freitas Mathias Paulo Cezar 
Published: April 2003

<i>Objective:</i> Pancreatic islets isolated from mice treated neonatally with monosodium <i>L</i>-glutamate (MSG) were used to study insulin secretion. <i>Material and Methods:</i> Total acetylcholinesterase (AchE) activity of tissue extract was measured as a cholinergic activity marker. Obesity recorded in 90-day-old MSG mice (OM) by Lee index reached 366.40 ± 1.70, compared to control mice (CM) 324.40 ± 1.10 (p < 0.0001). Glucose 5.6 m<i>M</i> induced insulin secretion of 36 ± 5 pg/15 min from islets of CM and 86 ± 13 from OM (p < 0.001). When glucose was raised to 16.7 m<i>M</i>, islets from OM secreted 1,271 ± 215 and 1,017 ± 112 pg/30 min to CM. AchE activity of pancreas from OM was 0.64 ± 0.02 nmol of substrate hydrolyzed/min/mg of tissue and 0.52 ± 0.01 to CM (p < 0.0001). Liver of obese animals also presented increase of AchE activity. <i>Results:</i> These indicate that OM insulin oversecretion in low glucose may be attributed, at least in part, to an enhancement of parasympathetic tonus.
Regulation of CD69 Expression on Eosinophil Precursors by Interferon-γ
Authors: Ochiai Kenichi , Kagami Masaru , Nakazawa Takuya , Sugiyama Takao , Sueishi Makoto , Ito Motohiro , Tomioka Hisao 
Published: April 2003

The purpose of this study was to determine whether interferon-γ (IFN-γ) induced CD69 expression by eosinophil precursors. Eosinophil precursors were induced from CD34+ cord blood cells using recombinant human interleukin-3 (IL-3) and interleukin-5 (IL-5). On day 14 of culture, cells constitutively expressed CD69 and the IFN-γ receptor (IFN-γR). Stimulation with IFN-γ for 24 h did not affect IFN-γR expression by the cells. On the other hand, IFN-γ significantly upregulated CD69 expression by the precursors after 24 h of incubation. A specific JAK2 inhibitor (AG-490) caused a concentration-dependent suppression of IFN-γ-induced CD69 expression by the precursors. In conclusion, these results indicate that IFN-γ induces CD69 expression by eosinophil precursors via the activation of JAK2.
Expression of Eotaxin by Normal Airway Epithelial Cells after Influenza Virus A Infection
Authors: Kawaguchi Mio , Kokubu Fumio , Kuga Hideki , Tomita Takeshi , Matsukura Satoshi , Kadokura Mitsutaka , Adachi Mitsuru 
Published: April 2003

<i>Background:</i> Viral infection is known to cause lung inflammatory disease, including bronchial asthma. The mechanisms of inflammatory cell accumulation into the airways after viral infection are not well understood. Eotaxin is a CC chemokine which is a potent and specific agonist for CC chemokine receptor 3 (CCR3). CCR3 is expressed on eosinophils, basophils and T lymphocytes. These cells are known to be key cells in the pathogenesis of asthma. Although it has recently been demonstrated that airway epithelial cells express eotaxin in vivo and in vitro, there are few data about its epxression in viral infection. We hypothesized that eotaxin may play an important role in attracting inflammatory cells to the airways after viral infection, and analyzed whether viral infection attracts eotaxin in bronchial epithelial cells in vitro. <i>Methods:</i> Human airway epithelial cells obtained from bronchial tissue at lobectomy for lung cancer were infected with influenza virus A (subtype H3N2). The cells and cultured media were collected 8, 24, and 48 h after infection. Eotaxin mRNA was analyzed with reverse transcriptase-polymerase chain reaction. Eotaxin protein levels in the culture media were analyzed by enzyme-linked immunosorbent assay. We also studied a blocking assay to analyze the intervention of proinflammatory cytokines in its production induced by influenza virus. <i>Results:</i> Eotaxin mRNA appeared to be expressed constitutively in uninfected cells but was expressed more clearly in infected cells. Eotaxin protein release into culture media significantly increased after infection. Anti-TNF-α and anti-IL-1β antibodies did not alter the eotaxin protein levels after viral infection. <i>Conclusions:</i> These results suggest that influenza virus A infection in airway epithelial cells activates the expression of eotaxin and that eotaxin may participate in the pathogenesis of airway inflammatory disease caused by viral infection, such as infectious type asthma.
An Optimal Condition of Bronchial Cell Proliferation Stimulated by Insulin-Like Growth Factor-I
Authors: Oyamada Hajime , Kayaba Hiroyuki , Kamada Yumiko , Kuwasaki Tomoe , Yamada Yoshiyuki , Kobayashi Yoshimi , Cui Chang-hao , Honda Kohei , Saito Norihiro , Chihara Junichi 
Published: April 2003

<i>Background:</i> It is known that growth hormones such as insulin-like growth factor-I (IGF-I) and several kinds of cytokines are involved in the regeneration process of injured epithelial cells in chronic respiratory inflammatory diseases such as bronchial asthma. Repetitive degeneration/regeneration processes of the airway epithelial layer is supposed to be responsible for the remodeling and irreversible organic changes of the airway in bronchial asthma. The purpose of this study is to establish a simple and reliable in vitro method for studying airway epithelial cell growth and proliferation using IGF-I. <i>Method:</i> By altering the number of cultured epithelial cells (strain NCI-H<sub>292</sub>), culture duration before stimulation with IGF-I, concentration of IGF-I, and duration of IGF-I stimulation, the optimum conditions for epithelial cell growth was determined. The epithelial cell growth was evaluated using [methyl-<sup>3</sup>H]thymidine uptake. <i>Result:</i> Among various culture conditions, the epithelial cells cultured at 1 × 10<sup>3</sup> cells/well for 24 h followed by 24 h of stimulation by 10<sup>–8</sup> <i>M</i> of IGF-I showed the highest growth. <i>Conclusion:</i> The method for the evaluation of epithelial cell growth established in this study requires a small number of cells and has no complicated procedure. This simple model enables us to investigate the effect of various substances on bronchial epithelial cell growth in the presence of IGF-I.
The Role of STAT1 in Activation of IL-3- and IL-5-Induced Eosinophils by Interferon Gamma
Authors: Ochiai K. , Otaka K. , Ito M. , Tomioka H. 
Published: April 2003

Tyrosine phosphorylation of STAT1α in eosinophils after IFN-γ stimulation has been shown, but the biological significance of eosinophil STAT1α activation in transmitting the signals through the IFN-γ receptor remains unknown. The purpose of this study is to determine whether STAT1 is involved in the regulation of eosinophils by IFN-γ-IFN-γ receptor interaction. rhIL-3- and rhIL-5-induced eosinophils from CD34+ cells of cord blood on day 28 of culture were used. The cells were washed and further incubated in IL-3- and IL-5-free medium for 48 h. The induced eosinophils constitutively expressed CD69 and lost this expression after a further 48-hour incubation without the cytokines. IFN-γ significantly upregulated CD69 expression on the 48-hour incubated cells. In inhibitory experiments on STAT1, a phosphorothioate oligo antisense DNA against STAT1α was added to IL-3- and IL-5-containing medium from day 15 to day 28 of culture. The oligo DNAs altered neither the expressions of myeloid cell marker CD9 and 13 nor the expression of IFN-γ receptor on the cells. The added STAT1α antisense, but not sense, DNA significantly reduced STAT1α mRNA expression in the cells. The STAT1 antisense also significantly inhibited IFN-γ-induced CD69 expression on the 48-hour incubated eosinophils. In conclusion, these results indicate that IFN-γ induces CD69 expression in the induced eosinophils through STAT1α, suggesting that STAT1α may play a significant role in eosinophil regulation by IFN-γ.
Intrauterine Treatment of Fetal Goitrous Hypothyroidism Controlled by Determination of Thyroid-Stimulating Hormone in Fetal Serum
Authors: Grüner Christian , Kollert Anja , Wildt Ludwig , Dörr Helmut G. , Beinder Ernst , Lang Norbert 
Published: April 2003

We report a rare case of fetal goitrous hypothyroidism complicated by polyhydramnios and preterm labor in a mother without thyroid gland pathology. The diagnosis was made in the 26th week by ultrasound and cordocentesis [TSH 170 µU/ml, free T<sub>4</sub> 0.2 ng/dl]. The therapeutic regime required repeated fetal blood sampling for determination of thyroid hormones. Five intra-amniotic administrations of 250 µg levothyroxine (LT4) weekyl were initiated. Because of the persisting goiter and the elevated level of TSH (128 µU/ml in 32 weeks) in the fetal serum the dosage had to be adjusted to 500 µg LT4 in the next five injections. TSH in fetal serum declined to 49.2 µU/ml in 36 weeks. Normal fetal growth and an uncomplicated course of pregnancy between the 27th and 37th week of gestation were observed. Monitoring of intrauterine therapy by determination of TSH in fetal serum may provide more reliable data than measuring TSH in amniotic fluid. A review of 15 cases of fetal goitrous hypothyroidism in the English literature is presented.
A Case of Massive Subchorionic Thrombohematoma Diagnosed by Ultrasonography and Magnetic Resonance Imaging
Authors: Kojima Kazuhisa , Suzuki Yoshikatsu , Makino Aiko , Murakami Isamu , Suzumori Kaoru 
Published: April 2003

Massive subchorionic thrombohematoma is uncommon but associated with a poor perinatal prognosis. Placental enlargement was detected in a 25-year-old Japanese primipara woman with fetal growth retardation and oligohydramnios at 23 weeks’ gestation. Ultrasonography (USG) showed an abnormal sonolucency within the placenta at 28 weeks’ gestation, but could not give an unequivocal differentiation from placental abnormalities such as hematomas, cysts and other tumors. Magnetic resonance imaging (MRI) pointed to a large hematoma in the subchorionic region. Simultaneously, the amniotic fluid was brownish colored. From these findings, it was possible to have prenatal diagnosis of massive subchorionic thrombohematoma. At 32 weeks’ gestation, the fetus died in utero and was stillborn 3 days later. Pathological findings for the placenta revealed a large hematoma diffused between the villous chorion and the chorionic plate, with wide necrosis of placental tissue, likely due to formation of multiple thrombi. The clinical and pathological findings were compatible with massive subchorionic thrombohematoma. MRI might be useful for the detection of massive subchorionic thrombohematoma and help its clinical management in combination with USG and pulse Doppler imaging.
Fetomaternal Transfusion and Pregnancy Outcome after Cordocentesis
Authors: Sikovanyecz János , Horváth Emese , Sallay Éva , Gellén János , Pál Attila , Szabó János 
Published: April 2003

<i>Objective:</i> To study the extent of fetomaternal transfusion and the outcome of pregnancy after cordocentesis. <i>Material and Methods:</i> 268 women underwent percutaneous fetal umbilical cord blood sampling for fetal karyotyping between 15 and 26 gestations of weeks. Complete follow-up was available in 221 (82.5%) of the cases. Cordocentesis was performed under continuous real-time ultrasound guidance. The duration of the procedure and the post-procedural bleeding time was counted in seconds. Fetomaternal transfusion was calculated by using the measurements of the maternal serum levels of α-fetoprotein before and after the procedure. The data were analyzed by Student’s t and multiple regression tests. <i>Results:</i> The maximum and mean amounts of fetomaternal transfusion were 1.067 and 0.061 ml, respectively. Twenty percent or more α-fetoprotein elevation was in 35.4% of the cases. Positive correlation was found between bleeding time after cordocentesis and fetomaternal transfusion (r = 0.174, p < 0.0129) as well as between the duration of the procedure (r = 0.165, p < 0.0171) and the amount of fetomaternal transfusion. Comparing the cordocentesis at the placental insertion site and at the free cord loop, a smaller amount of fetomaternal transfusion was observed (p < 0.0123) in the latter. Transplacental passage was associated with a higher amount of fetomaternal transfusion (p < 0.0067). No association was found between the extent of fetomaternal transfusion and the outcome of pregnancy. The fetal loss related to the cordocentesis was 0.50%. <i>Conclusions:</i> The extent of fetomaternal transfusion was influenced by the subsequent four parameters: procedural time, bleeding time, puncture site and transplacental penetration. The lack of the association between the degree of fetomaternal transfusion and the outcome of pregnancy, along with the low (0.50%) post-procedural fetal loss rate, suggest that cordocentesis is clinically a safe procedure.
Effects of Intra-Amniotic Meconium Exposure on the Fetal Rat: Development of a Pathogenic Model
Authors: Blackwell Sean C. , Hallak Mordechai , Hotra John W. , Hassan Sonia S. , Berry Stanley M. , Sorokin Yoram 
Published: April 2003

<i>Objective:</i> To develop an in vivo animal model for the study of the effects of intrauterine meconium exposure on the fetus. <i>Methods:</i> Timed pregnant Long-Evans rats were purchased on gestational day (GD) 12 and allowed to acclimate for at least 48 h prior to surgery. Laparotomy was performed and both uterine horns were exteriorized through the abdominal incision. A 26-gauge needle was used to inject either 0.1-cm<sup>3</sup> sterile normal saline or a 20% meconium suspension into each individual gestational sac. The uterus was returned to the abdomen and the incision was closed. On GD 21 (term = 21 days) a cesarean section was completed and the number and viability of fetuses in each horn were recorded. <i>Results:</i> A total of 14 animals were involved in this pilot study. One rat underwent sham surgery with only intra-amniotic saline injection and 13/15 fetuses survived to term. Two animals that underwent surgery on day 18 expired <24 h postinjection. Eleven maternal animals were injected on GD 20 and underwent cesarean delivery at term; survival rates for saline-injected animals were 71.2% compared to 66.2% for meconium-exposed fetuses. <i>Conclusion:</i> We have established an in vivo animal model that allows for the examination of the effects of prolonged intrauterine meconium exposure on the fetus.
Effects of Intrapartum Zidovudine Therapy on Fetal Heart Rate Parameters in Women with Human Immunodeficiency Virus Infection
Authors: Blackwell Sean C. , Sahai Anjali , Hassan Sonia S. , Treadwell Marjorie C. , Tomlinson Mark W. , Jones Theodore B. , Sorokin Yoram 
Published: April 2003

<i>Objective:</i> To determine the effects of intrapartum maternal administration of zidovudine on fetal heart rate (FHR) parameters in women infected with the human immunodeficiency virus (HIV). <i>Methods:</i> Term HIV-infected women who delivered at our institution (1995–1998) were identified by medical records coding. Sixty minutes of FHR tracing prior to zidovudine administration and 60 min of FHR tracing 2 h after initiation of therapy were reviewed by 3 perinatologists blinded to patient status. Data were compared with paired t tests; p < 0.05 was considered significant. Inter- and intra-observer FHR interpretation variation were calculated. <i>Results:</i> Ten patients met study criteria. Their demographic data included: Maternal age 26.5 ± 6.5 years, gestational age 38.9 ± 1.3 weeks, median parity 2 (range 0–3). Eighty percent were African American. There were no significant differences in FHR parameters after intravenous zidovudine therapy (pretreatment versus 2 h after treatment) with respect to FHR baseline (p = 0.2), FHR variability (p = 0.3), or the number of accelerations (p = 0.1). There was also no difference in the number of variable (moderate or severe), early, or prolonged decelerations following zidovudine infusion. <i>Conclusion:</i> Two hours of continuous intrapartum intravenous infusion of zidovudine does not alter any parameter of the FHR in the laboring HIV-infected gravida.
Development and Sequels of Intestinal Inflammation in Nematode-Infected Rats: Role of Mast Cells and Capsaicin-Sensitive Afferents
Authors: Gay Jerome , Fioramonti Jean , Garcia-Villar Rafael , Bueno Lionel 
Published: April 2003

<i>Objectives:</i> To determine whether intestinal mast cells and capsaicin-sensitive afferent nerves are involved in the development and sequels of <i>Nippostrongylus brasiliensis</i>-induced intestinal inflammation in rats. <i>Methods:</i> Two series of experiments were performed. In the first series, six groups of 8 rats were used to study the effects of mast cell stabilization by ketotifen. In the second series, six groups of 6 rats were used to study the effects of gut extrinsic sensory neuron depletion by capsaicin. For each series, four groups of rats were infected with <i>N. brasiliensis</i> and two groups were not infected. <i>Results:</i> Infection with <i>N. brasiliensis</i> resulted in an increase of myeloperoxidase (MPO) activity and mast cell numbers at day 12 postinfection; MPO returned to preinfection levels by day 35 while mast cell numbers remained elevated at that time. In ketotifen-treated infected rats, the increase of MPO at day 12 was less pronounced, but MPO activity remained elevated and mast cell numbers were increased at day 35. In capsaicin-treated infected rats, the MPO increase at day 12 was augmented, and MPO was still not returned to preinfection values by day 35; in contrast, the increase of mast cell numbers at days 12 and 35 was not modified by afferent nerve depletion. <i>Conclusion:</i> Mast cell stabilization decreased jejunal inflammation during the acute stage (day 12), but prolonged the inflammatory process until at least day 35 postinfection. The data also confirmed the protective role of gut extrinsic sensory neurons against intestinal inflammation in a model of nematode infection and revealed that these afferent nerves do not seem crucial for the development of nematode-induced hypermastocytosis.
Growth Hormone, Fatigue, Poor Sleep, and Disability in HIV Infection
Authors: Darko Denis F. , Mitler Merrill M. , Miller James C. 
Published: April 2003

Poor sleep, daytime fatigue, and loss of cognitive ability exist during all stages of HIV infection, worsening with disease progression. These symptoms contribute to disability and poor quality of life. Data from several research groups support a role of somnogenic inflammatory process peptides elevated in HIV infection, e.g. TNFα. Though the literature is in conflict regarding an effect of HIV infection on growth hormone (GH) secretion, GH axis dysregulation and treatment with GH may be important in HIV infection, e.g. in the wasting syndrome. It has long been known that GH varies with changes in sleep. The hypothesis tested in the current study was that the relationship between delta frequency (0.5–4.0 Hz) sleep EEG amplitude (square root of power from frequency analysis) and GH secretion would differ between HIV positive (HIV+) and HIV negative (HIV–) subjects. In 14 subjects (6 HIV+ and 8 HIV–, none with current or past AIDS-defining illness) a linear relationship change across the night’s sleep was found in the coupling between delta frequency sleep EEG amplitude and GH secretion. The phase coupling change was in opposite directions in HIV+ versus HIV– subjects. This difference supports the hypothesis that the brain-based coordination of sleep and sleep-related physiology deteriorates early in HIV infection, and that GH dysregulation may contribute to this sleep pathology.