bluuebunny/crossref_metadata_2025 huggingface dataset:
Eμ-BRD2 transgenic mice develop B-cell lymphoma and leukemia
Authors: Greenwald Rebecca J. , Tumang Joseph R. , Sinha Anupama , Currier Nicolas , Cardiff Robert D. , Rothstein Thomas L. , Faller Douglas V. , Denis Gerald V.
Published: October 2003

Transgenic mice with lymphoid-restricted overexpression of the double bromodomain protein bromodomain-containing 2 (Brd2) develop splenic B-cell lymphoma and, upon transplantation, B-cell leukemia with leukemic infiltrates in liver and lung. Brd2 is a nuclear-localized transcription factor kinase that is most closely related to TATA box binding protein–associated factor, 250 kDa (TAFII250) and the Drosophila developmental protein female sterile homeotic. Constitutive expression of BRD2 in the lymphoid compartment increases cyclin A transcription, “priming” transgenic B cells for proliferation. Mice stochastically develop an aggressive B-cell lymphoma with the features of B-1 cells, including CD5 and surface IgM expression. The B-cell lymphoma is monoclonal for immunoglobulin gene rearrangement and is phenotypically stable. The lymphoblasts are very large and express a transcriptome that is similar to human non-Hodgkin lymphomas. Both a wild-type BRD2 transgene and a kinase-null point mutant drive lymphomagenesis; therefore we propose that, rather than kinase activity, Brd2-mediated recruitment of E2 promoter binding factors (E2Fs) and a specific histone acetyltransferase to the cyclinA promoter by both types of transgene is a mechanistic basis for neoplasia. This report is the first to describe a transgenic mouse model for constitutive expression of a protein with more than one bromodomain.
Complete response to donor lymphocyte infusion in multiple myeloma is associated with antibody responses to highly expressed antigens
Authors: Bellucci Roberto , Wu Catherine J. , Chiaretti Sabina , Weller Edie , Davies Faith E. , Alyea Edwin P. , Dranoff Glenn , Anderson Kenneth C. , Munshi Nikhil C. , Ritz Jerome
Published: October 2003

The ability of donor lymphocyte infusions (DLIs) to induce complete responses (CRs) in patients with relapsed myeloma after allogeneic bone marrow transplantation (BMT) provides clear evidence of an effective graft-versus-myeloma (GVM) response. To identify target antigens of the GVM response, we screened a myeloma cDNA expression library with post-DLI serum from 4 patients with myeloma who achieved CR after DLI and 1 patient who was in CR before DLI. We identified a panel of 13 gene products reactive with post-DLI serum but negative with pre-DLI and pre-BMT serum. Antibodies to these proteins were not detected in the sera of 10 patients who underwent allogeneic BMT without DLI and 5 patients with acute graft-versus-host disease (GVHD). Minimal reactivity with these proteins was detected in the sera of 20 healthy donors and 20 patients with chronic GVHD. In contrast, 5 of these proteins were recognized by more than 1 myeloma DLI responder. Testing of serial serum samples showed an association between antibody response and time of best response after DLI. The expression of these genes was evaluated in primary myeloma cells and in normal plasma cells. This study demonstrates that the GVM response is associated with antibody responses to highly expressed myeloma-associated antigens.
Pharmacologically regulated Fas-mediated death of adoptively transferred T cells in a nonhuman primate model
Authors: Berger Carolina , Blau C. Anthony , Huang Meei-Li , Iuliucci John D. , Dalgarno David C. , Gaschet Joëlle , Heimfeld Shelly , Clackson Tim , Riddell Stanley R.
Published: October 2003

Conditional suicide genes derived from pathogens have been developed to confer drug sensitivity and enhance safety of cell therapy, but this approach is limited by immune responses to the transgene product. We examined a strategy to regulate survival of transferred cells based on induction of apoptosis through oligomerization of a modified human Fas receptor by a bivalent drug (AP1903). Three macaques (Macaca nemestrina) received autologous T cells retrovirally engineered to express a Fas suicide-construct (LV'VFas). High levels of transduced cells were present in blood following cell transfer, but LV'VFas+ cells declined rapidly after AP1903 administration. A small fraction of LV'VFas+ cells resisted elimination by AP1903, in part due to insufficient levels of transgene expression in resting T cells, because reactivation of these cells in vitro enhanced sensitivity to AP1903. An immune response to the transgene product was observed, but epitope mapping indicated the response was directed to discrete components of human LV'VFas that were variant with the corresponding macaque sequences. These data demonstrate that chemically induced dimerization can be used to regulate survival of adoptively transferred T cells in vivo.
Pleural drainage using central venous catheters
Authors: Singh Kulgit , Loo Shi , Bellomo Rinaldo
Published: October 2003

Introduction

The objective of the present study was to evaluate the use of a single lumen 16 G central venous catheter for the drainage of uncomplicated pleural effusions in intensive care unit patients.

Methods

A prospective observational study was performed in two intensive care units of university-affiliated hospitals. The study involved 10 intensive care unit patients with non-loculated large effusions. A 16 G central venous catheter was inserted at the bedside without ultrasound guidance using the Seldinger technique. The catheter was left in situ until radiological resolution of the effusion.

Results

Fifteen sets of data were obtained. The mean and standard deviation of the volumes drained at 1, 6 and 24 hours post catheter insertion were 454 ± 241 ml, 756 ± 403 ml and 1010 ± 469 ml, respectively. The largest volume drained in a single patient was 6030 ml over 11 days. The longest period for which the catheter remained in situ without evidence of infection was 14 days. There were no instances of pneumothorax, hemothorax, re-expansion pulmonary edema and catheter blockage/ disconnections.

Conclusions

The use of an indwelling 16 G central venous catheter is efficacious in draining uncomplicated large pleural effusions. It is well tolerated by patients and is associated with minimal complications. It has the potential to avoid repeated thoracentesis or the use of large-bore chest tubes.
Predictors of Occult Metastasis in Clinical Stage I Nonseminoma: A Systematic Review
Authors: Vergouwe Yvonne , Steyerberg Ewout W. , Eijkemans Marinus J.C. , Albers Peter , Habbema J. Dik F.
Published: October 2003

Purpose: Patients with clinical stage I nonseminomatous testicular germ cell tumor should ideally receive adjuvant therapy only when they are at high risk for occult metastasis. We aimed to quantify the importance of predictors for occult metastasis by performing a systematic review of the relevant literature. In addition, we reviewed published multivariable models and risk-adapted treatment policies.

Patients and Methods: We identified 23 publications between 1979 and 2001, reporting a total of 2,587 patients. Twenty-nine percent of the patients (759 of 2,587 patients) had occult metastases, which was diagnosed either at retroperitoneal lymph node dissection (n = 193) or during follow-up (n = 566). Odds ratios (OR) were pooled using meta-analysis techniques.

Results: The presence of vascular invasion of the primary tumor cells had the strongest effect (OR, 5.2; 95% CI, 4.0 to 6.8). Immunohistochemical staining of the primary tumor cells with the MIB-1 monoclonal antibody showing proliferative activity was a promising predictor (OR, 4.7; 95% CI, 2.0 to 11). Intermediate effects were found for embryonal carcinoma in the primary tumor (OR, 2.9; 95% CI, 2.0 to 4.4) and a high pathologic stage of the tumor (OR, 2.6; 95% CI, 1.8 to 3.8). Size of the primary tumor and age of the patient had weaker though also statistically significant associations with occult metastasis. Until now, multivariable models often included vascular invasion and embryonal carcinoma with one or two weaker predictors. None of the published risk-adapted treatment policies included MIB-1 staining.

Conclusion: Several strong predictors for occult metastasis were identified. A risk-adapted treatment policy should be developed that incorporates all relevant predictors so that adjuvant therapy is targeted better to those with occult metastases.
The Costs of Conducting Clinical Research
Authors: Emanuel Ezekiel J. , Schnipper Lowell E. , Kamin Deborah Y. , Levinson Jenifer , Lichter Allen S.
Published: October 2003

Purpose: Physicians frequently receive payment for enrolling subjects onto clinical trials. Some view these payments as conflicts of interest. Others contend that these payments are necessary reimbursements for conducting clinical research. We evaluated the clinical and nonclinical hours and costs associated with conducting a mock phase III clinical research trial.

Methods: We collected data from representatives of 21 clinical sites, on the numbers of hours associated with 13 activities necessary to the conduct of clinical research. The hours were based on enrolling 20 patients in a 12-month randomized placebo-controlled trial of a new chemotherapeutic agent. The outcome measures were disease progression and quality-of-life reports. These costs were evaluated for both government and pharmaceutical industry–sponsored trials.

Results: On average, 4,012 hours (range, 1,512 to 13,319 hours) were required for a government-sponsored trial, and 3,998 hours (range: 1735 to 15,699) were required for a pharmaceutical industry–sponsored trial involving 20 subjects with 17 office visits, or approximately 200 hours per subject. Thirty-two percent of the hours were devoted to nonclinical activities, such as institutional review board submission and completion of clinical reporting forms. On average, excluding overhead expenses, it cost slightly more than $6,094 (range, $2,098 to $19,285) per enrolled subject for an industry-sponsored trial, including $1,999 devoted to nonclinical costs.

Conclusion: Based on the results of our mock trial, the time required for nontreatment trial activities is considerable, and the associated costs are substantial.
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

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

Sulfonylureas 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

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

Human 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

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

The 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 < 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

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.
The amphioxus Hairy family: differential fate after duplication
Authors: Minguillón Carolina , Jiménez-Delgado Senda , Panopoulou Georgia , Garcia-Fernàndez Jordi
Published: October 2003

Vertebrate Hairy genes are highly pleiotropic and have been implicated in numerous functions, such as somitogenesis, neurogenesis and endocrine tissue development. In order to gain insight into the timing of acquisition of these roles by the Hairy subfamily, we have cloned and studied the expression pattern of the Hairy gene(s) in amphioxus. The cephalochordate amphioxus is widely believed to be the living invertebrate more closely related to vertebrates, the genome of which has not undergone the massive gene duplications that took place early during vertebrate evolution. Surprisingly,we have isolated eight Hairy genes from the `pre-duplicative' amphioxus genome. In situ hybridisation on amphioxus embryos showed that Hairy genes had experienced a process of subfunctionalisation that is predicted in the DDC model (for duplication-degeneration-complementation). Only the summation of four out of the eight Amphi-Hairy genes expression resembles the expression pattern of vertebrate Hairy genes, i.e. in the central nervous system,presomitic mesoderm, somites, notochord and gut. In addition, Amphi-Hairy genes expression suggest that amphioxus early somites are molecularly prefigured in an anteroposterior sequence in the dorsolateral wall of the archenteron, and the presence of a midbrain/hindbrain boundary. The expansion of the amphioxus Hairy subfamily request for caution when deducing the evolutionary history of a gene family in chordates based in the singularity of the amphioxus genome. Amphioxus may resemble the ancestor of the vertebrates,but it is not the ancestor, only its closest living relative, a privileged position that should not assume the freezing of its genome.
A contradictoryGLABRA3allele helps define gene interactions controlling trichome development inArabidopsis
Authors: Esch Jeffrey J. , Chen Margaret , Sanders Mark , Hillestad Matthew , Ndkium Sampson , Idelkope Brian , Neizer James , Marks M. David
Published: October 2003

Previously characterized Arabidopsis gl3 mutants have trichomes that are smaller, less branched and undergo fewer rounds of endoreplication than wild-type trichomes. A new gl3 mutant, called gl3-sst,has oddly shaped trichomes that over expand during early development, undergo more endoreduplication and that have a striking nuclear morphology. The mutant nuclei consist of many interconnected lobes; however, only a single set of polytene-like chromosomes reside in the mutant nuclei. The predicted gl3-sst polypeptide has a Leu to Phe substitution (codon 78) within a region responsible for protein-protein interaction. Yeast interaction assays comparing GL3 with gl3-sst proteins show that the mutant protein interaction with GL1 and TTG1 is decreased by 75% and 50%, respectively, but there is no difference in its interaction with TRY. Furthermore, TRY has the ability to prevent the GL1 GL3 interaction and the GL1 gl3-sst interaction is even more sensitive to TRY. Analysis of plants expressing functional GFP-tagged versions of GL1, GL3 and TRY show that the proteins are localized in trichome nuclei. These results have been used to model trichome initiation in terms of protein interactions and threshold levels of activator complex.
The role ofbuttonheadandSp1in the development of the ventral imaginal discs ofDrosophila
Authors: Estella Carlos , Rieckhof Gabrielle , Calleja Manuel , Morata Ginés
Published: October 2003

The related genes buttonhead (btd) and Drosophila Sp1 (the Drosophila homologue of the human SP1 gene)encode zinc-finger transcription factors known to play a developmental role in the formation of the Drosophila head segments and the mechanosensory larval organs. We report a novel function of btd and Sp1:they induce the formation and are required for the growth of the ventral imaginal discs. They act as activators of the headcase (hdc)and Distal-less (Dll) genes, which allocate the cells of the disc primordia. The requirement for btd and Sp1 persists during the development of ventral discs: inactivation by RNA interference results in a strong reduction of the size of legs and antennae. Ectopic expression of btd in the dorsal imaginal discs (eyes, wings and halteres) results in the formation of the corresponding ventral structures(antennae and legs). However, these structures are not patterned by the morphogenetic signals present in the dorsal discs; the cells expressing btd generate their own signalling system, including the establishment of a sharp boundary of engrailed expression, and the local activation of the wingless and decapentaplegic genes. Thus, the Btd product has the capacity to induce the activity of the entire genetic network necessary for ventral imaginal discs development. We propose that this property is a reflection of the initial function of the btd/Sp1 genes that consists of establishing the fate of the ventral disc primordia and determining their pattern and growth.
Posterior patterning genes and the identification of a unique body region in the brine shrimpArtemia franciscana
Authors: Copf Tijana , Rabet Nicolas , Celniker Susan E. , Averof Michalis
Published: October 2003

All arthropods share the same basic set of Hox genes, although the expression of these genes differs among divergent groups. In the brine shrimp Artemia franciscana, their expression is limited to the head,thoracic/trunk and genital segments, but is excluded from more posterior parts of the body which consist of six post-genital segments and the telson (bearing the anus). Nothing is currently known about the genes that specify the identity of these posterior structures. We examine the expression patterns of four candidate genes, Abdominal-B, caudal/Cdx, even-skipped/Evx and spalt, the homologues of which are known to play an important role in the specification of posterior structures in other animals. Abdominal-B is expressed in the genital segments of Artemia, but not in the post-genital segments at any developmental stage. The expression of caudal, even-skipped and spalt in the larval growth-zone suggests they may play a role in the generation of body segments (perhaps comparable with the role of gap and segmentation genes in insects), but not a direct role in defining the identity of post-genital segments. The expression of caudal at later stages suggests a role in the specification of anal structures. A PCR screen designed to isolate Hox genes expressed specifically in the posterior part of the body failed to identify any new Hox genes. We conclude that the post-genital segments of Artemia are not defined by any of the genes known to play a role in the specification of posterior segments in other arthropods. We argue that these segments constitute a unique body region that bears no obvious homology to previously characterised domains of Hox gene activity.
Meiotic germ cells antagonize mesonephric cell migration and testis cord formation in mouse gonads
Authors: Yao Humphrey H.-C. , DiNapoli Leo , Capel Blanche
Published: October 2003

The developmental fate of primordial germ cells in the mammalian gonad depends on their environment. In the XY gonad, Sry induces a cascade of molecular and cellular events leading to the organization of testis cords. Germ cells are sequestered inside testis cords by 12.5 dpc where they arrest in mitosis. If the testis pathway is not initiated, germ cells spontaneously enter meiosis by 13.5 dpc, and the gonad follows the ovarian fate. We have previously shown that some testis-specific events, such as mesonephric cell migration, can be experimentally induced into XX gonads prior to 12.5 dpc. However, after that time, XX gonads are resistant to the induction of cell migration. In current experiments, we provide evidence that this effect is dependent on XX germ cells rather than on XX somatic cells. We show that,although mesonephric cell migration cannot be induced into normal XX gonads at 14.5 dpc, it can be induced into XX gonads depleted of germ cells. We also show that when 14.5 dpc XX somatic cells are recombined with XY somatic cells,testis cord structures form normally; however, when XX germ cells are recombined with XY somatic cells, cord structures are disrupted. Sandwich culture experiments suggest that the inhibitory effect of XX germ cells is mediated through short-range interactions rather than through a long-range diffusible factor. The developmental stage at which XX germ cells show a disruptive effect on the male pathway is the stage at which meiosis is normally initiated, based on the immunodetection of meiotic markers. We suggest that at the stage when germ cells commit to meiosis, they reinforce ovarian fate by antagonizing the testis pathway.
To shatter the glass ceiling in healthcare management: who supports affirmative action and why?
Authors: Weil Peter A. , Mattis Mary C.
Published: October 2003

We examined the findings of a recent national survey of healthcare executives that showed 90% of women but only 53% of men favoured efforts to increase the proportion of women in senior healthcare management positions. Using the theories of relative deprivation and social identity, we tested hypotheses to suggest the background, work characteristics and attitudes about existing discriminatory practices in their own organizations that correlate with respondents' views about affirmative action for women. Some support is evidenced for the two theories and explanations are suggested to account for apparent anomalies.
Measuring progress in clinical governance: assessing the reliability and validity of the Clinical Governance Climate Questionnaire
Authors: Freeman T.
Published: October 2003

Despite a lack of conceptual clarity, the importance of cultural change to clinical governance is widely accepted. While generic measures of organizational performance, culture and climate are available, their relationship to clinical governance is unclear. Consequently, there is currently no valid and reliable measure of clinical governance climate. This study aimed to address the deficiency by reducing a pool of clinical governance climate indicators developed via previous qualitative research, describing a latent factor structure and assessing the internal consistency and external validity of the factor model. The resultant instrument, the Clinical Governance Climate Questionnaire (CGCQ), attained high internal consistency and external (discriminant and construct) validity in a study population of healthcare Trust staff. It consists of 60 items distributed across six sub-scales of clinical governance: planned and integrated quality improvement; proactive risk management; absence of unjust blame and punishment; working with colleagues; training and development; and organizational learning. The measure enables those charged with leading the clinical governance agenda in UK healthcare organizations to assess the progress of organizational development initiatives, highlighting areas requiring particular attention. They might also be of interest to those concerned about the negative unintended consequences of performance management.
From parallel policies to integrated practice: PMS and PCG/Ts
Authors: Walsh N. , Huntington J.
Published: October 2003

This paper describes findings from an exploratory study of attempts by primary care clinicians and managers to utilize two distinct Government policy initiatives (the establishment of Primary Care Groups [PCGs] and Trusts [PCTs] and the establishment of Primary Medical Services [PMS] pilots) to promote innovation in primary care. The study purposively selected three whole PCG-PMS sites and one PCT-PMS site. A range of different qualitative methods were used to collect data. Although the data suggest that attempts to integrate the two policies produced impressive corporate innovation by practices working together to benefit patients in whole localities, the four sites shared features that would not necessarily be present in all PCG/Ts. These include a strong history of inter-practice working, mutually supportive relationships between clinicians and managers, and a carefully designed, managed and inclusive change process led by credible clinicians. The study has implications for the continuing implementation of both policies, especially in view of the increasing size of PCTs.
Cyclic fluctuations in hospital bed occupancy in Roma (Italy): supply or demand driven?
Authors: Fusco D. , Saitto C. , Arcà M. , Ancona C. , Perucci C. A.
Published: October 2003

The objective of this study was to assess hospital bed occupancy both by planned and unplanned cases, and to assess how supply and demand affect bed occupancy. Data was obtained from the Lazio Hospital Information System (HIS) dataset on all hospital discharges from July 1998 to June 2001. Using Diagnosis Related Groups (DRG) as the reason for hospital stay, admissions were classified into four categories: 'planned stay', 'presumed planned stay', 'presumed unplanned stay', and 'unplanned stay'. Time series analysis of daily bed occupancy by category of stay was performed. Generalized Additive Models (GAMs) were used to asses the effect of weekdays and holidays on bed occupancy. Fluctuations in daily occupancy were observed in all categories of stay-in general, bed occupancy decreased over weekends, on national holidays, and during the major holiday season of August. In comparison with unplanned stays, the largest fluctuations were observed for planned stays while presumed planned and unplanned stays showed lesser fluctuations. It is possible to distinguish planned and unplanned hospital stays by using DRG grouping. Cyclic rigidities in the supply of services rather than the availability of beds or demand for beds seem to dictate hospital use in Roma so that restrictions in services hamper any reallocation of beds for 'planned stay' when demand for 'unplanned stay' beds declines.
Efavirenz: Enhancing the gold standard
Authors: Arribas J R
Published: October 2003

Efavirenz, the potent, once-daily non-nucleoside reverse transcriptase inhibitor, has been successfully used since 1996 and is still a cornerstone of antiretroviral therapy for use in treatment-naïve, treatment-experienced and difficult-to-treat patients. The efficacy of efavirenz in these patient groups has been established in many clinical and cohort studies. Furthermore, efavirenz is now available as a single 600 mg tablet, providing a simplified therapy and decreased pill burden, which may lead to improved adherence and treatment outcomes. Efavirenz has a well-characterized tolerability profile, the more notable side effects being mainly short-term and non-life-threatening, such as central nervous system disturbances and rash.
The power of simplicity
Authors: Lange J
Published: October 2003

Many years of use have confirmed didanosine to be among the most potent of nucleoside analogues. Didanosine's potency has been demonstrated in mono-, dual- and trip le-therapy. Didanosine is currently available as a once-daily, enteric-coated tablet, which has reduced gastrointestinal side effects compared with the older, buffered formulation. Once-daily dosing and good tolerability improve adherence, and hence also virological control. The prevalence of phenotypic nucleoside resistance to didanosine is low compared with that to zidovudine, lamivudine and abacavir. At least 6 nucleoside analogue mutations are needed in order for the virus to lose sensitivity to didanosine. Didanosine is effective in patients failing lamivudine therapy who have the M184V mutation. Enteric-coated didanosine is also an important component of early therapy, due to its potency and ease of administration.
The Assessing Patients’ Preferred Treatments (APPT-1) study
Authors:   the APPT-1Study Team, Moyle G
Published: October 2003

Limited information is currently available on patient preferences regarding administration frequency, tablet load and the trade-off between side effects and convenience in choosing antiretroviral regimens. A standardized questionnaire of persons with HIV infection across 5 large European countries was used to assess patient preference. 504 people with HIV participated. Participants showed a preference for once-daily therapy administration. However, several factors affected this preference.
Four policies to reduce HIV transmission through unsterile health care
Authors: Gisselquist David , Friedman Eric , Potterat John J , Minkin Stephen F , Brody Stuart
Published: October 2003

Residents of many developing countries face risks for themselves and their families to contract HIV and other bloodborne pathogens during unsterile health care. Helping people to understand and reduce these risks enlists their assistance to control the HIV/AIDS epidemic. To reduce HIV transmission through health care, we recommend four policies that international, foreign, and local public and private organizations can adopt and begin to implement even with little or no additional funds: (1) Educate the public about risks to contract HIV through unsterile health care. (2) Promote transparent practices for injections and other procedures that allow patients to see and know that care is safe (e.g., taking a new auto-disable syringe out of a sealed package and taking injecta from a single-dose vial). (3) Promote safe health care practices equally for clients and staff. (4) Establish a zero-tolerance policy for iatrogenic HIV infections, with publicly reported monitoring and investigations.
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.
HIV and AIDS in the People's Republic of China: a collaborative review
Authors: Portsmouth Simon , Stebbing Justin , Keyi Xu , Jianping Zhang , Guohua Pi
Published: October 2003

The number of individuals diagnosed with HIV in China has risen dramatically in the last two years coincident with increased awareness and an attitude change within government. UNAIDS has suggested that China could have 10 million HIV infected people by 2010. However, antiretroviral treatments and HIV testing are not yet widely available and infected individuals often live in remote areas. It is unlikely that cheaper, locally produced, generic antiretroviral formulations will be available in China in the near future. Consequently, alternative strategies to manage HIV infection are being considered including the use of hydroxyurea, chloroquine and traditional Chinese herbal medicines. It is recognized in China that prevention and educational strategies will need to be at the forefront of approaches to control this epidemic.