bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Effect on Tumor Response of Adding Sequential Preoperative Docetaxel to Preoperative Doxorubicin and Cyclophosphamide: Preliminary Results From National Surgical Adjuvant Breast and Bowel Project Protocol B-27
Authors: Bear Harry D. , Anderson Stewart , Brown Ann , Smith Roy , Mamounas Eleftherios P. , Fisher Bernard , Margolese Richard , Theoret Heather , Soran Atilla , Wickerham D. Lawrence , Wolmark Norman 
Published: October 2003

Purpose: The National Surgical Adjuvant Breast and Bowel Project Protocol B-27 was designed to determine the effect of adding docetaxel after four cycles of preoperative doxorubicin and cyclophosphamide (AC) on clinical and pathological response rates and on disease-free and overall survival of women with operable breast cancer. Patients and Methods: Women (N = 2,411) with operable primary breast cancer were randomly assigned to receive either four cycles of preoperative AC followed by surgery (group I), or four cycles of AC followed by four cycles of docetaxel, followed by surgery (group II), or four cycles of AC followed by surgery and then four cycles of docetaxel (group III). Clinical and pathologic tumor responses to preoperative therapy were assessed. Results: Mean tumor size (4.5 cm) and other key characteristics were evenly balanced among the three treatment arms. Grade 4 toxicity was observed in 10.3% of 2,400 patients during AC treatment, and in 23.4% of 1584 patients during docetaxel treatment. Compared to preoperative AC alone, preoperative AC followed by docetaxel increased the clinical complete response rate (40.1% v 63.6%; P < .001), the overall clinical response rate (85.5% v 90.7%; P < .001), the pathologic complete response rate (13.7% v 26.1%; P < .001), and the proportion of patients with negative nodes (50.8% v 58.2%; P < .001). Pathologic primary breast tumor response was a significant predictor of pathologic nodal status (P < .001). Conclusion: The addition of four cycles of preoperative docetaxel after four cycles of preoperative AC significantly increased clinical and pathologic response rates for operable breast cancer.
Functional Modification of Pituitary Somatotropes in the Aromatase Knockout Mouse and the Effect of Estrogen Replacement
Authors: Yan Ming , Jones Margaret E. E. , Hernandez Maria , Liu Dongling , Simpson Evan R. , Chen Chen 
Published: October 2003

Abstract
Available data on the influence of estradiol (E2) on GH levels remains controversial. A factor contributing to this uncertainty is a lack of knowledge of both E2 action on somatotropes as well as the molecular mechanisms involved. In this study we investigated gene expression implicated in GH secretion in somatotropes derived from female aromatase knockout (ArKO) mice. In these mice E2 production is blocked due to disruption of the Cyp19 gene encoding aromatase, the enzyme responsible for estrogen biosynthesis. The effect of E2 replacement was also studied by in vivo treatment of mice with E2 for 3 wk. It was demonstrated that somatotropes from ArKO mice had a low expression of GH, GH secretagogue receptor, GHRH receptor (GHRH-R), and pituitary-specific transcription factor (Pit-1). On the other hand, the somatotropes exhibited elevated expression of somatostatin receptors (sst1–5). Overall, these effects resulted in a reduction in GH secretion. E2 replacement increased GHRH-R, Pit-1, and GH mRNA levels to 185%, 193%, and 157% and reduced the levels of sst1, sst2, sst4, and sst5 mRNA expression in ArKO mice, respectively. E2 replacement did not affect the levels of pituitary estrogen (α and β) and androgen receptor mRNA expression. It is concluded that the expression of important genes involved in GH synthesis in somatotropes of the female ArKO mouse are functionally down-regulated, and such a down-regulation is reversed to normal levels by E2 replacement. The levels of GH secretagogue receptor, GHRH-R, and Pit-1 mRNA expression were also reduced, and sst1 and sst3 mRNA expression enhanced in aging ArKO and wild-type mice, resulting in a decrease in GH mRNA expression. It is suggested that aging is another important impact factor for the pituitary expression and regulation of GH mRNA in female mice.
Phosphatidylinositol 3-Kinase Activation Is Required for Sulfonylurea Stimulation of Glucose Transport in Rat Skeletal Muscle
Authors: Rodríguez Esther , Pulido Nieves , Romero Remedios , Arrieta Francisco , Panadero Aranzazu , Rovira Adela 
Published: October 2003

AbstractSulfonylureas are drugs widely used in the treatment of patients with type 2 diabetes mellitus. In addition to their pancreatic effect of stimulating insulin secretion, many studies suggest that sulfonylureas also have extrapancreatic actions. We have previously reported that gliclazide, a second-generation sulfonylurea, stimulates the glucose uptake by rat hindquarter skeletal muscle directly and immediately by promoting the translocation of glucose transporter 4 to the plasma membrane. The aim of our study was to approach the gliclazide intracellular signaling pathway. For this purpose, we incubated clamped and isolated soleus muscle from rat with gliclazide. The following results were obtained: 1) gliclazide stimulates insulin receptor substrate (IRS)-1-phosphatidylinositol 3 (PI3)-kinase-associated activity, and this activity is necessary for gliclazide-stimulated glucose transport; 2) gliclazide treatment produces a gradual translocation of the diacylglycerol (DAG)-dependent isoforms protein kinase C (PKC) α, θ, and ε from cytosolic to membrane fraction that is dependent on PI3-kinase and phospholipase C (PLC)-γ activation; and 3) PKC and PLC-γ activation is necessary for gliclazide-stimulated glucose transport. We propose a hypothetical signaling pathway by which gliclazide could stimulate IRS-1 that would allow its association with PI3-kinase, promoting its activation. PI3-kinase products could induce PLC-γ activation, whose hydrolytic activity could activate the DAG-dependent isoforms PKC α, θ, and ε.
Involvement of Cholesterol-Rich Lipid Rafts in Interleukin-6-Induced Neuroendocrine Differentiation of LNCaP Prostate Cancer Cells
Authors: Kim Jayoung , Adam Rosalyn M. , Solomon Keith R. , Freeman Michael R. 
Published: October 2003

Abstract
IL-6 is an inflammatory cytokine that has been linked to aggressive prostate cancer (PCa). Previous studies have demonstrated that IL-6 can enhance the differentiation of PCa cells toward a neuroendocrine (NE) phenotype, a possible indicator of hormone-refractory disease. In this report, we present evidence that the mechanism of IL-6-stimulated NE differentiation employs a detergent-resistant (lipid raft) membrane compartment for signal transduction in LNCaP PCa cells. Signal transducer and activator of transcription (STAT)3, a mediator of IL-6 signaling, was rapidly phosphorylated and translocated to the nucleus in LNCaP cells treated with IL-6. Both processes were inhibited by filipin, a cholesterol-binding compound that disrupts plasma membrane lipid rafts. Isolation of Triton X-100-insoluble raft fractions from LNCaP cells by discontinuous sucrose gradient centrifugation demonstrated that the 80-kDa IL-6 receptor localized almost exclusively to the raft compartment. Although STAT3 was located predominantly in the Triton X-100-soluble subcellular fraction in exponentially growing cells, abundant phosphorylated STAT3 was detected in the raft fraction after stimulation with IL-6. Increases in expression of the NE marker, neuron-specific enolase, and neuron-specific enolase promoter activity after IL-6 treatment were reduced after membrane rafts were disrupted by filipin treatment. LNCaP cells expressed the raft-resident proteins flotillin-2 and Giα2, but notably not caveolins, the predominant structural protein present in caveolar membrane rafts in many tissues and tumor cells. These results are the first to define a role for lipid raft membrane microdomains in signal transduction mechanisms capable of promoting the NE phenotype in PCa cells, and they demonstrate that the raft compartment is capable of mediating such signals in the absence of caveolins. Our results also suggest a mechanistic role for membrane cholesterol in cell signaling events relevant to PCa progression.
The Role of Cyclic-ADP-Ribose-Signaling Pathway in Oxytocin-Induced Ca2+ Transients in Human Myometrium Cells
Authors: Barata Hosana , Thompson Michael , Zielinska Weronika , Han Young S. , Mantilla Carlos B. , Prakash Yedatore S. , Feitoza Simone , Sieck Gary , Chini Eduardo N. 
Published: October 2003

AbstractHuman myometrial contraction plays a fundamental role in labor. Dysfunction of uterine contraction is an important cause of labor progression failure. Although the mechanisms controlling uterine contraction are not completely understood, intracellular Ca2+ mobilization plays an important role during uterine contraction. Several mechanisms of intracellular Ca2+ mobilization are present in smooth muscle, but in the human uterus, only 1,4,5-trisphosphate-induced Ca2+ release has been studied extensively. Ryanodine receptor channels are present in myometrium. We determined the role of the cyclic ADP-ribose (cADPR)-signaling pathway in oxytocin-induced intracellular Ca2+ [(Ca2+)i] transients in human myometrial cells. We found that oxytocin-induced Ca2+ transient is dependent on several sources of Ca2+, including extracellular Ca2+ and intracellular Ca2+ stores. In addition, we found that both the 1,4,5-trisphosphate- and the cADPR-induced Ca2+ releasing systems are important for the induction of [Ca2+]i transients by oxytocin in human myometrial cells. Furthermore, we investigated TNFα regulation of oxytocin-induced [Ca2+]i transients, CD38 cyclase activity, and CD38 expression in human myometrial cells. We found that oxytocin-induced [Ca2+]i transients were significantly increased by 50 ng/ml TNF. Similarly, CD38 mRNA levels, CD38 expression, and cyclase activity were increased by TNFα, thus increasing cADPR levels. We propose that a complex interaction between multiple signaling pathways is important for the development of intracellular Ca2+ transients induced by oxytocin and that TNFα may contribute for the myometrium preparation for labor by regulating the cADPR-signaling pathway. The observation that the cADPR-signaling pathway is important for the development of intracellular Ca2+ transients in human myometrial cells raises the possibility that this signaling pathway could serve as a target for the development of new therapeutic strategies for abnormal myometrial contraction observed during pregnancy.
Cathepsin L Gene Expression and Promoter Activation in Rodent Granulosa Cells
Authors: Sriraman Venkataraman , Richards JoAnne S. 
Published: October 2003

Abstract
The cysteine protease cathepsin L exhibits hormone-regulated expression during ovulation. In situ hybridization analyses of immature and pregnant mare serum gonadotropin-treated mouse and rat ovaries showed that cathepsin L expression in granulosa cells of small, growing follicles increased in periovulatory follicles after human chorionic gonadotropin stimulation. In the rat ovary, cathepsin L was also expressed in follicles with signs of atresia. To determine the molecular mechanisms that mediate the diverse regulation of this gene in granulosa cells, rat cathepsin L promoter-reporter constructs were analyzed by transient transfection assays in rat granulosa cells and EMSAs. A construct containing the transcriptional start site and −244 bp of upstream promoter sequence (−244/+33 bp) exhibited inducibility by forskolin, the phorbol ester phorbol myristate acetate, and an additive effect of both. Within this region, three functional specificity protein 1 (Sp1) sites, an overlapping early growth response protein-1 site, and a cAMP regulatory element-binding protein site were identified. Single or double mutants of the above-mentioned sites did not alter forskolin/phorbol myristate acetate inducibility of the promoter. Mutation of all three Sp1/specificity protein 3 (Sp3) sites, which also mutated the early growth response protein-1 site, reduced the promoter activation. Mutation of the cAMP regulatory element-binding protein site in the triple Sp1 mutant construct completely blocked the inducibility of the promoter. When these same constructs were transfected into MCF-7 human breast cancer cells or were cotransfected with an Sp1 expression vector in Drosophila SL2 cells, similar results were obtained. Collectively, the data document that three Sp1/specificity protein 3 binding GC-rich regions and a functional cAMP regulatory element constitute an important transcriptional regulatory complex for expression of the cathepsin L gene in rat granulosa cells.
Localization of ISG15 and Conjugated Proteins in Bovine Endometrium Using Immunohistochemistry and Electron Microscopy
Authors: Austin Kathy J. , Carr Amy L. , Pru James K. , Hearne Carol E. , George Evee L. , Belden E. Lee , Hansen Thomas R. 
Published: October 2003

AbstractThe interferon-stimulated gene ISG15, a ubiquitin homolog, becomes conjugated to and regulates uterine proteins in response to conceptus-derived interferon-τ on d 18 of pregnancy. It was hypothesized here that cellular localization of ISG15 within endometrial cells might provide insight regarding function. Uteri were collected from cows (∼21-d estrous cycle) on d 17–21/0 of the estrous cycle and pregnancy and d 23, 45, and 50 of pregnancy. Intracellular ISG15 and its conjugates were present on d 17 of pregnancy, peaked to highest levels from d 18 to 23 and then declined to low but detectable levels by d 45 (P &lt; 0.05) based on Western blotting. ISG15 and its conjugates were not detected on d 50 of pregnancy or during the estrous cycle. Immunohistochemistry revealed that ISG15 was localized throughout the endometrium on d 18–23, with heaviest staining in the sublumenal stratum compactum and the glandular epithelium throughout the stratum spongiosum. By d 45 and 50, ISG15 was lightly stained only in the stratum compactum immediately beneath the lumenal epithelium. Using transmission electron microscopy and immunogold labeling, ISG15 was specifically localized to organelles and compartments of endometrial epithelial cells and stromal cells: nucleus, perinuclear space, cytosol, mitochondria, rough endoplasmic reticulum, and cell membrane. This specific localization in epithelial and stromal cells led to the conclusion that ISG15 has diverse intracellular functions. The sustained presence of conjugated ISG15 through d 50 of pregnancy might reflect stabilization of conjugated proteins in response to implantation and the development of the placenta.
Cytochrome P450 2D Catalyze Steroid 21-Hydroxylation in the Brain
Authors: Kishimoto Wataru , Hiroi Toyoko , Shiraishi Masakazu , Osada Mayuko , Imaoka Susumu , Kominami Shiro , Igarashi Takashi , Funae Yoshihiko 
Published: October 2003

Abstract
mRNA of cytochrome P450 21-hydroxylase (P450c21) is expressed in the brain, but little is known about the enzymatic properties of P450c21 in the brain. In the present study, we showed, by using various recombinant cytochrome P450 (CYP)2D enzymes and anti-CYP2D4- or P450c21-specific antibodies, that rat brain microsomal steroid 21-hydroxylation is catalyzed not by P450c21, but by CYP2D isoforms. Rat CYP2D4 and human CYP2D6, which are the predominant CYP2D isoforms in the brain, possess 21-hydroxylation activity for both progesterone and 17α-hydroxyprogesterone. In rat brain microsomes, these activities were not inhibited by anti-P450c21 antibodies, but they were effectively inhibited by the CYP2D-specific chemical inhibitor quinidine and by anti-CYP2D4 antibodies. mRNA and protein of CYP2D4 were expressed throughout the brain, especially in cerebellum, striatum, pons, and medulla oblongata, whereas the mRNA and protein levels of P450c21 were extremely low or undetectable. These results support the idea that CYP2D4, not P450c21, works as steroid 21-hydroxylase in the brain. Allopregnanolone, a representative γ-aminobutyric acid receptor modulator, was also hydroxylated at the C-21 position by recombinant CYP2D4 and CYP2D6. Rat brain microsomal allopregnanolone 21-hydroxylation was inhibited by fluoxetine with an IC50 value of 2 μm, suggesting the possibility that the brain CYP2D isoforms regulate levels of neurosteroids such as allopregnanolone, and that this regulation is modified by central nervous system-active drugs such as fluoxetine.
Screening for genital chlamydia infection: DNA amplification techniques should be the test of choice
Authors: Harindra V , Underhill G , Tobin J M 
Published: October 2003

Our objective was to compare the sensitivities for the detection of Chlamydia trachomatis, of the ligase chain reaction (LCR) on first voided urine (FVU) specimens and enzyme immunoassay (EIA) on pooled endocervical/endourethral swabs from women and endourethral swabs from men. Men and women taking part in the UK chlamydia screening pilot were tested for chlamydia using LCR on a FVU. Patients attending genitourinary medicine clinics also had cervical and/or urethral swabs taken for chlamydia testing by EIA. In women, EIA on pooled swabs detected 575 of the 785 chlamydia positives and in men, EIA detected 209 of 351 positives. The sensitivity of EIA was 73% and 60% in women and men respectively. By using the EIA test, therefore, 27-40% of patients infected with chlamydia will be given a false negative result. We propose that it is unethical to use non-molecular testing in the future.
Continued high-risk sexual behaviour among HIV-positive people in Wales
Authors: Browning Mary R , Evans Meirion R , Rees Clive M 
Published: October 2003

The study objective was to investigate self-reported sexual practices among HIV-positive people in Wales in order to estimate the extent of unprotected sex and to develop services that assist positive people reduce the risk of sexual spread of HIV. A cross-sectional postal questionnaire survey was conducted between February and July 1999 at all open-access HIV outpatient facilities in Wales. All HIV-positive people attending during the study period were invited to enrol in the study. Data were obtained on 104/141 (74%) enrolled patients including 70 (67%) homo/bisexual men. Most (79/104, 76%) were sexually active and 42% (33/79) had casual partners. One in six (13/79, 16.5%) sexually active people reported unprotected high-risk sex with HIVnegative or unknown status partners, the majority (9/13, 69%) of whom were male homosexuals. It is important that health professionals specifically address issues of sexual behaviour with HIV positive people during consultation. A renewed safer sex campaign targeting men who have sex with men is urgently needed.
HIV-1 Plasma RNA level and CD4 cell count in a large urban HIV-1-infected patient population from 1997 to 2000
Authors: Shan Hua , Piwowar-Manning Estelle , Thompson Richard E , Brooks Jackson J 
Published: October 2003

We evaluated the change in plasma HIV-1 RNA level and CD4 cell counts in an HIV-1-infected population between 1997 and 2000. Both the mean and the median values of plasma HIV-1 RNA level decreased with time with the exception of 1998. The mean and medians for CD4 cell count appear to be fairly stable. While the percentage of patients with plasma HIV-1 RNA level <400 copies/mL increased from 29.12% in 1997 to 41.31% in 2000, the percentage of patients with plasma HIV-1 RNA level >100,000 copies/mL remained fairly constant. The availability of HAART had impacted the level of plasma HIV-1 RNA, although many patients still have plasma HIV-1 RNA level >100,000 copies/mL. Most patients with plasma HIV-1 RNA level >400 copies/mL in 1997 still have plasma HIV-1 RNA level >400 copies/mL in 2000. These laboratory findings, however, do not necessarily mean that there had been a lack of clinical benefit.
Prevalence of asymptomatic chlamydial urethritis in military recruits in the Celje region, Slovenia
Authors: Skaza A , Grsković B , Plestina S , Bozina N , Potocnik M , Waugh M A 
Published: October 2003

The aim of this research was to evaluate the prevalence of asymptomatic chlamydial urethritis in military recruits in the Celje region (population 300,000), Slovenia. A first-void urine specimen was tested for Chlamydia trachomatis using the polymerase chain reaction assay. The research was supported by a questionnaire to obtain information on sexual behaviour of the participants. In the cross-sectional study from 1999 to 2001, 1272 asymptomatic recruits were included. None had received antibiotics in the previous two weeks. The mean age was 19.9 years. At the time of their first sexual experience the mean age was 16.6 years, whereas the age of their female sexual partners was 17.1 years. During their first sexual intercourse 77% of recruits used contraception (condom, diaphragm, contraceptive pill), most of those a condom (86%). The prevalence of asymptomatic chlamydial urethritis was 2.6% (95% confidence interval: 1.7 to 3.5). The mean age of those infected was 19.8 years. At the time of their first sexual experience the mean age was 16.2 years, whereas the age of their female sexual partners was 16.9 years. During their first sexual intercourse 57% of infected subjects used protection, half of which was a condom. Those who never or only occasionally used condoms were at a greater risk of being infected with C. trachomatis (adjusted odds ratio 2.04).