bluuebunny/crossref_metadata_2025 huggingface dataset:
Chim3 confers survival advantage to CD4+ T cells upon HIV-1 infection by preventing HIV-1 DNA integration and HIV-1–induced G2 cell-cycle delay
Authors: Porcellini Simona , Gubinelli Francesco , Alberici Luca , Piovani Bianca Maria , Rizzardi Gian-Paolo , Bovolenta Chiara
Published: March 2010

The long-term expression and the ability of a therapeutic gene to confer survival advantage to transduced cells are mandatory requirements for successful anti-HIV gene therapy. In this context, we developed lentiviral vectors (LVs) expressing the F12–viral infectivity factor (Vif) derivative Chim3. We recently showed that Chim3 inhibits HIV-1 replication in primary cells by both blocking the accumulation of retrotranscripts, independently of either human APOBEC3G (hA3G) or Vif, and by preserving the antiviral function of hA3G. These results were predictive of long-lasting survival of Chim3+ cells after HIV-1 infection. Furthermore, Vif, like Vpr, deregulates cell-cycle progression by inducing a delay in G2 phase. Thus, the aim of this study was to investigate the role of Chim3 on both cell survival and cell-cycle regulation after HIV-1 infection. Here, we provide evidence that infected Chim3+ T cells prevail over either mock- or empty-LV engineered cells, show reduced G2 accumulation, and, as a consequence, ultimately extend their lifespan. Based on these findings, Chim3 rightly belongs to the most efficacious class of antiviral genes. In conclusion, Chim3 usage in anti-HIV gene therapy based on hematopoietic stem cell (HSC) modification has to be considered as a promising therapeutic intervention to eventually cope with HIV-1 infection.
Allogeneic transplantation improves the overall and progression-free survival of Hodgkin lymphoma patients relapsing after autologous transplantation: a retrospective study based on the time of HLA typing and donor availability
Authors: Sarina Barbara , Castagna Luca , Farina Lucia , Patriarca Francesca , Benedetti Fabio , Carella Angelo M. , Falda Michele , Guidi Stefano , Ciceri Fabio , Bonini Alessandro , Ferrari Samantha , Malagola Michele , Morello Enrico , Milone Giuseppe , Bruno Benedetto , Mordini Nicola , Viviani Simonetta , Levis Alessandro , Giordano Laura , Santoro Armando , Corradini Paolo ,   for Gruppo Italiano Trapianto di Midollo Osseo
Published: March 2010

Hodgkin lymphoma relapsing after autologous transplantation (autoSCT) has a dismal outcome. Allogeneic transplantation (alloSCT) using reduced intensity conditioning (RIC) is a salvage option, but its effectiveness is still unclear. To evaluate the role of RIC alloSCT, we designed a retrospective study based on the commitment of attending physicians to perform a salvage alloSCT; thus, only Hodgkin lymphoma patients having human leukocyte antigen-typing immediately after the failed autoSCT were included. Of 185 patients, 122 found an identical sibling (55%), a matched unrelated (32%) or a haploidentical sibling (13%) donor; 63 patients did not find any donor. Clinical features of both groups did not differ. Two-year progression-free (PFS) and overall survival (OS) were better in the donor group (39.3% vs 14.2%, and 66% vs 42%, respectively, P < .001) with a median follow-up of 48 months. In multivariable analysis, having a donor was significant for better PFS and OS (P < .001). Patients allografted in complete remission showed a better PFS and OS. This is the largest study comparing RIC alloSCT versus conventional treatment after a failed autoSCT, indicating a survival benefit for patients having a donor.
Role of prostaglandin E receptor subtypes EP2 and EP4 in autocrine and paracrine functions of vascular endothelial growth factor in the inner ear
Authors: Hori Ryusuke , Nakagawa Takayuki , Yamamoto Norio , Hamaguchi Kiyomi , Ito Juichi
Published: March 2010

Background

The physiological effects of prostaglandin E1 (PGE1) and prostaglandin E2 (PGE2) are mediated by the prostaglandin E receptor subtypes EP1, EP2, EP3, and EP4, and the respective agonists have been purified. PGE1 and PGE2 can increase the production of vascular endothelial growth factor (VEGF), particularly through EP2 and EP4. The biological effects of VEGF are mediated by the phosphotyrosine kinase receptors fms-related tyrosine kinase-1 (Flt-1) and fetal liver kinase-1 (Flk-1). Here we examined the effects of EP2 and EP4 agonists on the production of VEGF proteins andVEGFmessenger RNAs (mRNAs) in the inner ear, using an enzyme-linked immunosorbent assay and the real-time quantitative reverse transcription-polymerase chain reaction, respectively. We also examined the localization of EP2, VEGF, Flt-1, and Flk-1 in the cochlea by immunohistochemistry.

Results

The expression of EP2 occurred in the cochlea, and the local application of an EP2 or EP4 agonist increased VEGF protein andVEGFmRNA levels in the inner ear. Furthermore, the intensity of the VEGF immunoreactivity in the spiral ganglion appeared to be increased by the local EP2 or EP4 agonist treatment. Immunoreactivity for Flt-1, and Flk-1 was found in the cochlear sensory epithelium, spiral ganglion, spiral ligament, and stria vascularis.

Conclusions

These findings demonstrate that EP2 and EP4 agonists stimulate VEGF production in the inner ear, particularly in the spiral ganglions. Moreover, the Flt-1 and Flk-1 expression observed in the present study suggests that VEGF has autocrine and paracrine actions in the cochlea. Thus, EP2 and EP4 might be involved in the mechanisms underlying the therapeutic effects of PGE1 on acute sensorineural hearing loss via VEGF production.
Variants of the Matrix Metalloproteinase-2 but not the Matrix Metalloproteinase-9 genes significantly influence functional outcome after stroke
Authors: Manso Helena , Krug Tiago , Sobral João , Albergaria Isabel , Gaspar Gisela , Ferro José M , Oliveira Sofia A , Vicente Astrid M
Published: March 2010

Background

Multiple lines of evidence suggest that genetic factors contribute to stroke recovery. The matrix metalloproteinases -2 (MMP-2) and -9 (MMP-9) are modulators of extracellular matrix components, with important regulatory functions in the Central Nervous System (CNS). Shortly after stroke, MMP-2 and MMP-9 have mainly damaging effects for brain tissue. However, MMPs also have a beneficial activity in angiogenesis and neurovascular remodelling during the delayed neuroinflammatory response phase, thus possibly contributing to stroke functional recovery.

Methods

In the present study, the role of MMP-2 and MMP-9 genetic variants in stroke recovery was investigated in 546 stroke patients. Functional outcome was assessed three months after a stroke episode using the modified Rankin Scale (mRS), and patients were classified in two groups: good recovery (mRS ≤ 1) or poor recovery (mRS>1). Haplotype tagging single nucleotide polymorphisms (SNPs) in the MMP-2 (N = 21) and MMP-9 (N = 4) genes were genotyped and tested for association with stroke outcome, adjusting for significant non-genetic clinical variables.

Results

Six SNPs in the MMP-2 gene were significantly associated with stroke outcome (0.0018<P < 0.0415), two of which survived the Bonferroni correction for multiple testing. In the subset of ischemic stroke patients, association of five of these SNPs remained positive (0.0042<P < 0.0306). No significant associations were found for the MMP-9 gene.

Conclusions

The results presented strongly indicate that MMP-2 genetic variants are an important mediator of functional outcome after stroke.
Use of the internet as a source of health information by Spanish adolescents
Authors: Jiménez-Pernett Jaime , de Labry-Lima Antontio Olry , Bermúdez-Tamayo Clara , García-Gutiérrez Jose Francisco , del Carmen Salcedo-Sánchez Maria
Published: March 2010

Background

The Internet is a fundamental part of the daily life of adolescents, they consider it as a safe and confidential source of information on health matters. The aims is to describe the experience of Spanish adolescents searching for health information on the Internet.

Methods

A cross-sectional study of 811 school-age adolescents in Granada was carried out. An adapted and piloted questionnaire was used which was controlled by trained personnel. Sociodemographic and health variables were included together with those concerning the conditions governing access to and use of information and communication technologies (ICT).

Results

811 adolescents were surveyed (99.38% response rate), mean age was 17 years old. Of these, 88% used the Internet; 57.5% used it on a daily or weekly basis and 38.7% used it occasionally. Nearly half the sample group (55.7%) stated that they used the Internet to search for health-related information. The main problems reported in the search for e-health were the ignorance of good web pages (54.8%) and the lack of confidence or search skills (23.2%).

Conclusions

In conclusion, it seems plausible to claim that websites designed and managed by health services should have a predominant position among interventions specifically addressed to young people.
Control of mandibular incisors with the combined Herbst and completely customized lingual appliance - a pilot study
Authors: Wiechmann Dirk , Schwestka-Polly Rainer , Pancherz Hans , Hohoff Ariane
Published: March 2010

Background

The traditional Herbst appliance induces mandibular incisor proclination independent of the anchorage system used. The dental effects of the Herbst appliance as an element of a completely customized lingual orthodontic (LO) appliance (Incognito, 3 M) has not been analyzed yet and the aim of this paper was to measure the effect of mandibular incisor proclination using this Herbst-LO device.

Methods

Retrospective study. Inclusion criteria: a) Class II ≥ 5 mm molar relationship; b) Herbst appliance ≥ 9 months in situ; and c) finished active treatment. Incisor position was measured on digital models before treatment, on the digital target setup, and on digital models obtained at the day of debonding. All measurements were performed by one investigator.

Results

Twelve patients (8 females, 4 males) out of 632 cases treated with a lingual appliance were included in the study. The measurement error computed with Dahlberg's formula was 0.2°. Seven cases had planned (target setup) mandibular incisor uprighting (ccr), and five cases had proclination (clockwise rotation). There was no statistical difference (p > 0.05) between planned incisor rotations of the target setup and achieved incisor rotations at the day of debonding. The overall mean difference was 2.2° ± 1.0°.

Conclusions

The Incognito-Herbst combination is the first Herbst device with full control over mandibular incisor movement. Using this system, anchorage loss or anchorage gain is independent of the Herbst treatment. It depends only on the planned tooth position of the individual target setup.
Revision hip replacement for recurrent Hydatid disease of the pelvis: a case report and review of the literature
Authors: Neelapala Venkata SS , Chandrasekar Coonoor R , Grimer Robert J
Published: March 2010

A case of a large recurrent hydatid cyst involving the right ilium and right hip treated with excision of the cyst, Total hip replacement and revision of the acetabular component with a Tripolar articulation for cyst recurrence and acetabular component loosening is presented along with a review of the relevant literature. To our knowledge there is no reported case of Total Hip replacement and revision for hydatid disease involving the bony pelvis.
Phase 2 study of canfosfamide in combination with pegylated liposomal doxorubicin in platinum and paclitaxel refractory or resistant epithelial ovarian cancer
Authors: Kavanagh John J , Levenback Charles F , Ramirez Pedro T , Wolf Judith L , Moore Carla L , Jones Marsha R , Meng Lisa , Brown Gail L , Bast Robert C
Published: March 2010

Background

Canfosfamide is a novel glutathione analog activated by glutathione S-transferase P1-1. This study evaluated the safety and efficacy of canfosfamide in combination with pegylated liposomal doxorubicin (PLD) in patients with platinum resistant ovarian cancer. Patients with platinum resistant ovarian carcinoma and measurable disease received canfosfamide at 960 mg/m2in combination with PLD at 50 mg/m2, intravenously day 1 in every 28 day cycles until tumor progression or unacceptable toxicities. The primary endpoints were objective response rate (ORR) and progression-free survival (PFS).

Results

Canfosfamide plus PLD combination therapy was administered at 960/50 mg/m2, respectively. Thirty-nine patients received a median number of 4 cycles (range 1.0-18.0). The ORR was 27.8% (95% CI, 14.2-45.2) with a disease stabilization rate of 80.6% (95% CI, 64.0-91.8) in the evaluable population. The CA-125 marker responses correlated with the radiological findings of complete response or partial response. The median PFS was 6.0 months (95% CI, 4.2-7.9) and median survival was 17.8 months. The combination was well tolerated. Myelosuppression was managed with dose reductions and growth factor support. Grade 3 febrile neutropenia was observed in 2 patients (5.1%). Non-hematologic adverse events occurred at the expected frequency and grade for each drug alone, with no unexpected or cumulative toxicities.

Conclusions

Canfosfamide in combination with PLD is well tolerated and active in platinum and paclitaxel refractory or resistant ovarian cancer. A randomized phase 3 study was conducted based on this supportive phase 2 study.

Trial Registration

This study was registered at www.clinicaltrials.gov: NCT00052065.
Effect of Prior Bilateral Oophorectomy on the Presentation of Breast Cancer in BRCA1 and BRCA2 Mutation Carriers
Authors: Metcalfe Kelly A , Foulkes William D , Lynch Henry T , Ghadirian Parviz , Tung Nadine , Olivotto Ivo A , Warner Ellen , Olopade Olufunmilayo , Eisen Andrea , Weber Barbara , McLennan Jane , Sun Ping , Narod Steven A
Published: March 2010

Purpose

To compare the presentation of invasive breast cancer in BRCA1 and BRCA2 mutation carriers with and without prior bilateral oophorectomy.

Patients and methods

Women with a BRCA1 or BRCA2 mutation with the diagnosis of invasive breast cancer were identified from ten cancer genetics clinics. The medical history, medical treatment records and pathology reports for the breast cancers were reviewed. Information was abstracted from medical charts, including history (and date) of oophorectomy, date of breast cancer diagnosis, stage of disease, and pathologic characteristics of the breast cancer. Women with prior bilateral oophorectomy were matched by age, year of diagnosis, and mutation with one or more women who had two intact ovaries at the time of breast cancer diagnosis. Characteristics of the breast tumours were compared between the two groups.

Results

Women with prior bilateral oophorectomy presented with smaller tumours on average compared to women without prior oophorectomy (mean size 1.50 cm vs. 1.95 cm; p = 0.01). Additionally, although not statistically significant, women with intact ovaries were more likely to have high-grade tumour (70% vs. 54%: p = 0.10) and to have positive lymph nodes (34% vs. 18%; p = 0.11) compared to women with prior bilateral oophorectomy.

Conclusions

Bilateral oophorectomy prior to breast cancer appears to favourably influence the biological presentation of breast cancer in BRCA1 and BRCA2 mutation carriers.
Clinical, Molecular and Geographical Features of Hereditary Breast/Ovarian Cancer in Latvia
Authors: Gardovskis Andris , Irmejs Arvids , Miklasevics Edvins , Borosenko Viktors , Bitina Marianna , Melbarde-Gorkusa Inga , Vanags Andrejs , Kurzawski Grzegorz , Suchy Janina , Górski Bohdan , Gardovskis Janis
Published: March 2010

Introduction

The aim of the study is to evaluate the incidence and phenotype-genotype characteristics of hereditary breast and ovarian cancer syndromes in Latvia in order to develop the basis of clinical management for patients and their relatives affected by this syndrome.

Materials and methods

In 2002-2004 in two Latvian oncology hospitals (Liepãja Oncology Hospital and Daugavpils Oncology Hospital) cancer family histories were collected from 287 consecutive patients with breast and ovarian cancer. In all cases, when it was possible to obtain the blood sample, DNA testing for founder mutations in the BRCA1 gene was performed.

Results

Among 287 family cancer histories analysed in 8 (2.8%) cases criteria of hereditary breast cancer (HBC) were fulfilled and in 5 (1.7%) cases hereditary breast and ovarian cancer (HBOC) was diagnosed. In 50 (17.4%) cases we have suspicion of hereditary breast cancer (HBC susp.) and in 8 (2.8%) cases - suspicion of hereditary breast and ovarian cancer (HBOC susp.). We have one (0.3%) case with hereditary ovarian cancer (HOC). DNA testing of founder mutations in the BRCA1 gene (exon 20 (5382 insC) exon 5 (300T/G), exon 11, 17 (4153delA)) for 178/287 (62%) patients was performed. In 9/287 (4.9%) cases we found a mutation in the BRCA1 gene. 4 mutations were detected in exon 11, 17 (4153delA) and 4 mutations in exon 20 (5382 insC) and 1 in exon 5.

Conclusions

Existing pedigree/clinical data suggest that in Latvia the clinical frequency of hereditary breast and ovarian cancer is around 5% of consecutive breast and ovarian cancer patients and suspicion of the syndrome is observed in another 20% of cases. Frequency of BRCA1 founder mutations is 5% of all consecutive breast and ovarian cancers. Considerable geographical differences in the clinical and molecular frequency of hereditary breast ovarian cancer have been observed in Latvia.
Cysteinyl-leukotriene type 1 receptors transduce a critical signal for the up-regulation of eosinophilopoiesis by interleukin-13 and eotaxin in murine bone marrow
Authors: Queto Tulio , Gaspar-Elsas Maria I , Masid-de-Brito Daniela , Vasconcelos Zilton F M , Ferraris Fausto K , Penido Carmen , Cunha Fernando Q , Kanaoka Yoshihide , Lam Bing K , Xavier-Elsas Pedro
Published: March 2010

Novel role of eotaxin and IL-13 in regulating bone-marrow eosinophilopoiesis, and a critical role for CysLT1R receptors in transducing eotaxin/IL-13 signals.

IL-13 and eotaxin play important, inter-related roles in asthma models. In the lungs, CysLT, produced by the 5-LO-LTC4S pathway, mediate some local responses to IL-13 and eotaxin; in bone marrow, CysLT enhance IL-5-dependent eosinophil differentiation. We examined the effects of IL-13 and eotaxin on eosinophil differentiation. Semi-solid or liquid cultures were established from murine bone marrow with GM-CSF or IL-5, respectively, and the effects of IL-13, eotaxin, or CysLT on eosinophil colony formation and on eosinophil differentiation in liquid culture were evaluated, in the absence or presence of: a) the 5-LO inhibitor zileuton, the FLAP inhibitor MK886, or the CysLT1R antagonists, montelukast and MK571; b) mutations that inactivate 5-LO, LTC4S, or CysLT1R; and c) neutralizing mAb against eotaxin and its CCR3 receptor. Both cytokines enhanced GM-CSF-dependent eosinophil colony formation and IL-5-stimulated eosinophil differentiation. Although IL-13 did not induce eotaxin production, its effects were abolished by anti-eotaxin and anti-CCR3 antibodies, suggesting up-regulation by IL-13 of responses to endogenous eotaxin. Anti-CCR3 blocked eotaxin completely. The effects of both cytokines were prevented by zileuton, MK886, montelukast, and MK571, as well as by inactivation of the genes coding for 5-LO, LTC4S, and CysLT1R. In the absence of either cytokine, these treatments or mutations had no effect. These findings provide evidence for: a) a novel role of eotaxin and IL-13 in regulating eosinophilopoiesis; and b) a role for CysLTRs in bone marrow cells in transducing cytokine regulatory signals.
Phospholipase D2 Mediates Acute Aldosterone Secretion in Response to Angiotensin II in Adrenal Glomerulosa Cells
Authors: Qin Haixia , Frohman Michael A. , Bollag Wendy B.
Published: March 2010

In primary bovine adrenal glomerulosa cells, the signaling enzyme phospholipase D (PLD) is suggested to mediate priming, the enhancement of aldosterone secretion after pretreatment with and removal of angiotensin II (AngII), via the formation of persistently elevated diacylglycerol (DAG). To further explore PLD’s role in priming, glomerulosa cells were pretreated with an exogenous bacterial PLD. Using this approach, phosphatidic acid (PA) is generated on the outer, rather than the inner, leaflet of the plasma membrane. Although PA is not readily internalized, the PA is nonetheless rapidly hydrolyzed by cell-surface PA phosphatases to DAG, which efficiently flips to the inner leaflet and accesses the cell interior. Pretreatment with bacterial PLD resulted in priming upon subsequent AngII exposure, supporting a role of DAG in this process, because the increase in DAG persisted after exogenous PLD removal. To determine the PLD isoform mediating aldosterone secretion, and presumably priming, primary glomerulosa cells were infected with adenoviruses expressing GFP, PLD1, PLD2, or lipase-inactive mutants. Overexpressed PLD2 increased aldosterone secretion by approximately 3-fold over the GFP-infected control under basal conditions, with a significant enhancement to about 16-fold over the basal value upon AngII stimulation. PLD activity was also increased basally and upon stimulation with AngII. In contrast, PLD1 overexpression had little effect on aldosterone secretion, despite the fact that PLD activity was enhanced. In both cases, the lipase-inactive PLD mutants showed essentially no effect on PLD activity or aldosterone secretion. Our results suggest that PLD2 is the isoform that mediates aldosterone secretion and likely priming.
Exendin-4 Prevents c-Jun N-Terminal Protein Kinase Activation by Tumor Necrosis Factor-α (TNFα) and Inhibits TNFα-Induced Apoptosis in Insulin-Secreting Cells
Authors: Natalicchio Annalisa , De Stefano Francesca , Orlando Maura Roberta , Melchiorre Mariangela , Leonardini Anna , Cignarelli Angelo , Labarbuta Rossella , Marchetti Piero , Perrini Sebastio , Laviola Luigi , Giorgino Francesco
Published: March 2010

Glucagon-like peptide-1 and its analogs may preserve pancreatic β-cell mass by promoting resistance to cytokine-mediated apoptosis. The mechanisms of TNFα-induced apoptosis and of its inhibition by exendin-4 were investigated in insulin-secreting cells. INS-1 and MIN6 insulinoma cells were exposed to 20 ng/ml TNFα, with or without pretreatment with 10 nm exendin-4. Treatment with TNFα increased c-Jun N-terminal protein kinase (JNK) phosphorylation 2-fold, reduced inhibitor-κBα (IκBα) protein content by 50%, induced opposite changes in caspase-3 and Bcl-2 protein content, and increased cellular apoptosis. Moreover, exposure to TNFα resulted in increased serine phosphorylation of both insulin receptor substrate (IRS)-1 and IRS-2 and reduced basal and insulin-induced Akt phosphorylation. However, in the presence of a JNK inhibitor, TNFα-induced apoptosis was diminished and serine phosphorylation of IRS proteins was prevented. When cells were pretreated with exendin-4, TNFα-induced JNK and IRS-1/2 serine phosphorylation was markedly reduced, Akt phosphorylation was increased, caspase-3 and Bcl-2 protein levels were restored to normal, and TNFα-induced apoptosis was inhibited by 50%. This was associated with a 2-fold increase in IRS-2 expression levels. A similar ability of exendin-4 to prevent TNFα-induced JNK phosphorylation was found in isolated pancreatic human islets. The inhibitory effect of exendin-4 on TNFα-induced JNK phosphorylation was abrogated in the presence of the protein kinase A inhibitor H89. In conclusion, JNK activation mediates TNFα-induced apoptosis and impairment of the IRS/Akt signaling pathway in insulin-secreting cells. By inhibiting JNK phosphorylation in a PKA-dependent manner, exendin-4 counteracts TNFα-mediated apoptosis and reverses the inhibitory events in the IRS/Akt pathway, resulting in promotion of cell survival.
The Positive Effects of Growth Hormone-Releasing Peptide-6 on Weight Gain and Fat Mass Accrual Depend on the Insulin/Glucose Status
Authors: Granado Miriam , García-Cáceres Cristina , Frago Laura M. , Argente Jesús , Chowen Julie A.
Published: March 2010

Ghrelin and GH secretagogues, including GH-releasing peptide (GHRP)-6, stimulate food intake and adiposity. Because insulin modulates the hypothalamic response to GH secretagogues and acts synergistically with ghrelin on lipogenesis in vitro, we analyzed whether insulin plays a role in the metabolic effects of GHRP-6 in vivo. Streptozotocin-induced diabetic rats received saline, GHRP-6, insulin, or insulin plus GHRP-6 once daily for 8 wk. Rats receiving saline suffered hyperglycemia, hyperphagia, polydipsia, and weight loss. Insulin, but not GHRP-6, improved these parameters (P < 0.001 for all), as well as the diabetes-induced increase in hypothalamic mRNA levels of neuropeptide Y and agouti-related peptide and decrease in proopiomelanocortin. Cocaine amphetamine-related transcript mRNA levels were also reduced in diabetic rats, with GHRP-6 inducing a further decrease (P < 0.03) and insulin an increase. Diabetic rats receiving insulin plus GHRP-6 gained more weight and had increased epididymal fat mass and serum leptin levels compared with all other groups (P < 0.001). In epididymal adipose tissue, diabetic rats injected with saline had smaller adipocytes (P < 0.001), decreased fatty acid synthase (FAS; P < 0.001), and glucose transporter-4 (P < 0.001) and increased hormone sensitive lipase (P < 0.001) and proliferator-activated receptor-γ mRNA levels (P < 0.01). Insulin normalized these parameters to control values. GHRP-6 treatment increased FAS and glucose transporter-4 gene expression and potentiated insulin’s effect on epididymal fat mass, adipocyte size (P < 0.001), FAS (P < 0.001), and glucose transporter-4 (P < 0.05). In conclusion, GHRP-6 and insulin exert an additive effect on weight gain and visceral fat mass accrual in diabetic rats, indicating that some of GHRP-6’s metabolic effects depend on the insulin/glucose status.
Expression of E-Cadherin and N-Cadherin in Perinatal Hamster Ovary: Possible Involvement in Primordial Follicle Formation and Regulation by Follicle-Stimulating Hormone
Authors: Wang Cheng , Roy Shyamal K.
Published: March 2010

We examined the expression and hormonal regulation of E-cadherin (CDH1) and N-cadherin (CDH2) with respect to primordial follicle formation. Hamster Cdh1 and Cdh2 cDNA and amino acid sequences were more than 90% similar to those of the mouse, rat, and human. Although CDH1 expression remained exclusively in the oocytes during neonatal ovary development, CDH2 expression shifted from the oocytes to granulosa cells of primordial follicles on postnatal day (P)8. Subsequently, strong CDH2 expression was restricted to granulosa cells of growing follicles. Cdh2 mRNA levels in the ovary decreased from embryonic d 13 through P10 with a transient increase on P7, which was the day before the appearance of primordial follicles. Cdh1 mRNA levels decreased from embryonic d 13 through P3 and then showed a transient increase on P8, coinciding with the formation of primordial follicles. CDH1 and CDH2 expression were consistent with that of mRNA. Neutralization of FSH in utero impaired primordial follicle formation with an associated decrease in Cdh2 mRNA and CDH2, but an increase in Cdh1 mRNA and CDH1 expression. The altered expression was reversed by equine chorionic gonadotropin treatment on P1. Whereas a CDH2 antibody significantly reduced the formation of primordial and primary follicles in vitro, a CDH1 antibody had the opposite effect. This is the first evidence to suggest that primordial follicle formation requires a differential spatiotemporal expression and action of CDH1 and CDH2. Further, FSH regulation of primordial follicle formation may involve the action of CDH1 and CDH2.
Cadmium Promotes Breast Cancer Cell Proliferation by Potentiating the Interaction between ERα and c-Jun
Authors: Siewit Christina L. , Gengler Bridget , Vegas Esera , Puckett Rachel , Louie Maggie C.
Published: March 2010

Cadmium is an environmental contaminant that enters the body through diet or cigarette smoke. It affects multiple cellular processes, including cell proliferation, differentiation, and apoptosis. Recently, cadmium has been shown to function as an endocrine disruptor, to stimulate estrogen receptor α (ERα) activity and promote uterine and mammary gland growth in mice. Although cadmium exposure has been associated with the development of breast cancer, the mechanism of action of cadmium remains unclear. To address this deficit, we examined the effects of cadmium treatment on breast cancer cells. We found that ERα is required for both cadmium-induced cell growth and modulation of gene expression. We also determined that ERα translocates to the nucleus in response to cadmium exposure. Additionally, we provide evidence that cadmium potentiates the interaction between ERα and c-Jun and enhances recruitment of this transcription factor complex to the proximal promoters of cyclin D1 and c-myc, thus increasing their expression. This study provides a mechanistic link between cadmium exposure and ERα and demonstrates that cadmium plays an important role in the promotion of breast cancer.
Research Resource: Expression Profiling Reveals Unexpected Targets and Functions of the Human Steroid Receptor RNA Activator (SRA) Gene
Authors: Foulds Charles E. , Tsimelzon Anna , Long Weiwen , Le Andrew , Tsai Sophia Y. , Tsai Ming-Jer , O'Malley Bert W.
Published: March 2010

The human steroid receptor RNA activator (SRA) gene encodes both noncoding RNAs (ncRNAs) and protein-generating isoforms. In reporter assays, SRA ncRNA enhances nuclear receptor and myogenic differentiation 1 (MyoD)-mediated transcription but also participates in specific corepressor complexes, serving as a distinct scaffold. That SRA RNA levels might affect some biological functions, such as proliferation, apoptosis, steroidogenesis, and myogenesis, has been reported. However, the breadth of endogenous target genes that might be regulated by SRA RNAs remains largely unknown. To address this, we depleted SRA RNA in two human cancer cell lines with small interfering RNAs and then assayed for changes in gene expression by microarray analyses. The majority of significantly changed genes were reduced upon SRA knockdown, implicating SRA RNAs as endogenous coactivators. Unexpectedly, only a small subset of direct estrogen receptor-α target genes was affected in estradiol-treated MCF-7 cells. Eight bona fide SRA downstream target genes were identified (SLC2A3, SLC2A12, CCL20, TGFB2, DIO2, TMEM65, TBL1X, and TMPRSS2), representing entirely novel SRA targets, except for TMPRSS2. These data suggest unanticipated roles for SRA in glucose uptake, cellular signaling, T3 hormone generation, and invasion/metastasis. SRA depletion in MDA-MB-231 cells reduced invasiveness and expression of some genes critical for this process. Consistent with the knockdown data, overexpressed SRA ncRNA coactivates certain target promoters and may enhance the activity of some coregulatory proteins. This study is a valuable resource because it represents the first genome-wide analysis of a mammalian RNA coregulator.
Chemical enhancement of torsinA function in cell and animal models of torsion dystonia
Authors: Cao Songsong , Hewett Jeffrey W. , Yokoi Fumiaki , Lu Jun , Buckley Amber Clark , Burdette Alexander J. , Chen Pan , Nery Flavia C. , Li Yuqing , Breakefield Xandra O. , Caldwell Guy A. , Caldwell Kim A.
Published: March 2010

Movement disorders represent a significant societal burden for which therapeutic options are limited and focused on treating disease symptomality. Early-onset torsion dystonia (EOTD) is one such disorder characterized by sustained and involuntary muscle contractions that frequently cause repetitive movements or abnormal postures. Transmitted in an autosomal dominant manner with reduced penetrance, EOTD is caused in most cases by the deletion of a glutamic acid (ΔE) in the DYT1 (also known as TOR1A) gene product, torsinA. Although some patients respond well to anticholingerics, therapy is primarily limited to either neurosurgery or chemodenervation. As mutant torsinA (ΔE) expression results in decreased torsinA function, therapeutic strategies directed toward enhancement of wild-type (WT) torsinA activity in patients who are heterozygous for mutant DYT1 may restore normal cellular functionality. Here, we report results from the first-ever screen for candidate small molecule therapeutics for EOTD, using multiple activity-based readouts for torsinA function in Caenorhabditis elegans, subsequent validation in human DYT1 patient fibroblasts, and behavioral rescue in a mouse model of DYT1 dystonia. We exploited the nematode to rapidly discern chemical effectors of torsinA and identified two classes of antibiotics, quinolones and aminopenicillins, which enhance WT torsinA activity in two separate in vivo assays. Representative molecules were assayed in EOTD patient fibroblasts for improvements in torsinA-dependent secretory function, which was improved significantly by ampicillin. Furthermore, a behavioral defect associated with an EOTD mouse knock-in model was also rescued following administration of ampicillin. These combined data indicate that specific small molecules that enhance torsinA activity represent a promising new approach toward therapeutic development for EOTD, and potentially for other diseases involving the processing of mutant proteins.
Valuing Multifactor Real Options Using an Implied Binomial Tree
Authors: Wang Tianyang , Dyer James S.
Published: March 2010

This paper proposes an approach for solving a multifactor real options problem by approximating the underlying stochastic process with an implied binomial tree. The implied binomial tree is constructed to be consistent with simulated market information. By simulating European option prices as artificial market information, we apply the implied binomial tree method for real options valuation when the options are contingent on the value of market uncertainties that are not traded assets. Compared to the discrete approximations suggested in the current literature, this method offers a more flexible distribution assumption for project values and therefore provides an alternative approach to estimating the value of high-dimensional real options. For risk managers, it serves as a capital budgeting method for projects with managerial flexibility.
Return on Roller Coasters: A Model to Guide Investments in Theme Park Attractions
Authors: van Oest Rutger D. , van Heerde Harald J. , Dekimpe Marnik G.
Published: March 2010

Despite the economic significance of the theme park industry and the huge investments needed to set up new attractions, no marketing models exist to guide these investment decisions. This study addresses this gap in the literature by estimating a response model for theme park attendance. The model not only determines the contribution of each attraction to attendance, but also how this contribution is distributed within and across years. The model accommodates saturation effects, which imply that the impact of a new attraction is smaller if similar attractions are already present. It also captures reinforcement effects, meaning that a new attraction may reinforce the drawing power of similar extant attractions, especially when these were introduced recently. The model is calibrated on 25 years of weekly attendance data from the Efteling, a leading European theme park. Our return on investment calculations show that it is more profitable to invest in multiple smaller attractions than in one big one. This finding is in remarkable contrast with the current “arms race” in the industry. Furthermore, even though thrill rides tend to be more effective than theme rides, there are conditions under which one should consider to switch to the latter.
The Impact of Customer Community Participation on Customer Behaviors: An Empirical Investigation
Authors: Algesheimer René , Borle Sharad , Dholakia Utpal M. , Singh Siddharth S.
Published: March 2010

Many firms increasingly offer community venues to their customers to facilitate social interactions amongst them. Prior studies have shown that community participants have high engagement and loyalty toward the firm and provide useful feedback and referrals. However, it is not clear whether community participants are the firm's “fans” to begin with and self-select themselves into the community, or whether community participation leads to increased relational customer behaviors. In the current research, we employ data from a field experiment to help answer this question. The data come from a year-long study conducted by eBay Germany, and they reveal that a simple e-mail invitation significantly increased customer participation in the firm's community. Results also show that community participation had mixed effects on customers' likelihoods of participating in buying and selling behaviors. Community participation did not translate into increased behaviors, as would be commonly expected. Although there is no impact of participation on the number of bids placed or the revenue earned, there is a negative impact of participation on the number of listings and the amount spent. Together, these results suggest that the community participants become more selective and efficient sellers, and they also become more conservative in their spending on the items for which they bid. The results also show that customer community marketing programs may be targeted to a broader set of the firm's customers than just the fans.
Positive Effects of Negative Publicity: When Negative Reviews Increase Sales
Authors: Berger Jonah , Sorensen Alan T. , Rasmussen Scott J.
Published: March 2010

Can negative information about a product increase sales, and if so, when? Although popular wisdom suggests that “any publicity is good publicity,” prior research has demonstrated only downsides to negative press. Negative reviews or word of mouth, for example, have been found to hurt product evaluation and sales. Using a combination of econometric analysis and experimental methods, we unify these perspectives to delineate contexts under which negative publicity about a product will have positive versus negative effects. Specifically, we argue that negative publicity can increase purchase likelihood and sales by increasing product awareness. Consequently, negative publicity should have differential effects on established versus unknown products. Three studies support this perspective. Whereas a negative review in the New York Times hurt sales of books by well-known authors, for example, it increased sales of books that had lower prior awareness. The studies further underscore the importance of a gap between publicity and purchase occasion and the mediating role of increased awareness in these effects.
Growing Two-Sided Networks by Advertising the User Base: A Field Experiment
Authors: Tucker Catherine , Zhang Juanjuan
Published: March 2010

Two-sided exchange networks (such as eBay.com) often advertise their number of users, presumably to encourage further participation. However, these networks differ markedly on how they advertise their user base. Some highlight the number of sellers, some emphasize the number of buyers, and others disclose both. We use field experiment data from a business-to-business website to examine the efficacy of these different display formats. Before each potential seller posted a listing, the website randomized whether to display the number of buyers and/or sellers, and if so, how many buyers and/or sellers to claim. We find that when information about both buyers and sellers is displayed, a large number of sellers deters further seller listings. However, this deterrence effect disappears when only the number of sellers is presented. Similarly, a large number of buyers is more likely to attract new listings when it is displayed together with the number of sellers. These results suggest the presence of indirect network externalities, whereby a seller prefers markets with many other sellers because they help attract more buyers.
Long-Term Aspirin and Clopidogrel Response Evaluated by Light Transmission Aggregometry, VerifyNow, and Thrombelastography in Patients Undergoing Percutaneous Coronary Intervention
Authors: Madsen Esben Hjorth , Saw Jacqueline , Kristensen Søren Risom , Schmidt Erik Berg , Pittendreigh Cheryl , Maurer-Spurej Elisabeth
Published: March 2010

Background: A reduced response to aspirin and clopidogrel predicts ischemic events, but reliable tests are needed to identify low responders. We compared 3 platelet-function tests during long-term dual treatment with aspirin and clopidogrel.

Methods: Patients who underwent a percutaneous coronary intervention and were receiving a combination of 325 mg/day aspirin and 75 mg/day clopidogrel were followed for 1 year. Blood was sampled 5 times during this period for 3 tests: light transmission aggregometry (LTA) assay, with 5.0 μmol/L ADP or 1.0 mmol/L arachidonic acid (AA) used as an agonist; VerifyNow™ assay, with the P2Y12 or aspirin cartridge (Accumetrics); and thrombelastography (TEG), stimulated by 2.0 μmol/L ADP or 1.0 mmol/L AA.

Results: Twenty-six of 33 patients completed all scheduled visits. A low response to clopidogrel was found in a few patients at variable frequencies and at different visits, depending on the method and criteria used. We found a moderate correlation between the LTA (ADP) and VerifyNow (P2Y12 cartridge) results, but the TEG (ADP) results correlated poorly with the LTA and VerifyNow results. A low response to aspirin was found with the VerifyNow (aspirin cartridge) and TEG (AA) methods on 6 and 2 occasions, respectively, but not with the LTA (AA) method, except for 1 occasion caused by probable noncompliance.

Conclusions: Detecting a low response to clopidogrel depends largely on the method used. Which method best predicts ischemic events remains uncertain. A low response to aspirin is rare with AA-dependent methods used at the chosen cutoffs. In some patients, the response to clopidogrel or aspirin may be classified differently at different times, even with the same method.
Aneuploidy Detection in Mixed DNA Samples by Methylation-Sensitive Amplification and Microarray Analysis
Authors: Brown Lucia , Brown Gregory , Vacek Pamela , Brown Stephen
Published: March 2010

Background: Cell-free fetal nucleic acid, believed to be derived from the placenta/trophoblast, is present in the plasma of pregnant women; however, its use for predictive genetic testing has been severely limited because the circulating fetal DNA is present in a small quantity and mixed with a much larger quantity of maternal DNA. Methods for detecting fetal aneuploidy from the cell-free fetal DNA in plasma are highly sought after, but proposed methods must take into account the small quantity and highly contaminated nature of the available fetal DNA.

Methods: We developed a method for methylation-sensitive amplification of DNA suitable for use with small (approximately 1 ng) samples. We used this method in conjunction with 2-color microarray analysis with a custom-made array to investigate whether relative amplification, and hence relative methylation, could be evaluated for a large number of genomic loci.

Results: Microarray assessment of genomic methylation accurately predicted the degree of methylation measured with bisulfite-conversion PCR and confirmed that DNA from first-trimester trophoblast was generally hypomethylated compared with whole-blood DNA. With a series of 3 samples in which 1 ng of DNA from a trisomic first trimester placenta was mixed with 9 ng of chromosomally normal peripheral blood DNA, we observed that the microarray signal associated with the trisomic chromosome was significantly different from that of the other chromosomes (P < 0.001).

Conclusions: This method has potential to be used for noninvasive detection of fetal aneuploidy.
Genetic polymorphisms of CYP1A2, CYP3A4, CYP3A5, pregnane/steroid X receptor and constitutive androstane receptor in 207 healthy Spanish volunteers
Authors: Oliver Paloma , Lubomirov Rubin , Carcas Antonio
Published: March 2010

Background:Variability of cytochrome P450 (CYP) in humans is largely related to the pharmacological and toxicological effects of drugs and chemicals. Identification of single nucleotide polymorphisms (SNPs) could be important for knowing their involvement in many drugs metabolism. The goal of this study was to analyze the genotype frequency of 10 SNPs related to mirtazapine metabolism [CYP3A4*17, CYP3A4*18, CYP3A5*3A, CYP1A2*1F, pregnane/steroid X receptor (PXR) (rs3814055, rs38114057, rs3814058) and constitutive androstane receptor (CAR) (rs4073054, rs2307424, rs2502815)].

Methods:The study was carried out in 207 healthy Spanish volunteers that had participated in phase I clinical trials. Other studies were performed: Hardy-Weinberg equilibrium, haplotype estimation and linkage disequilibrium.

Results:No mutation related to CYP3A4*17 and CYP3A4*18 was found. Therefore, we analyzed data for the other eight SNPs. Allele frequencies were in equilibrium with the Hardy-Weinberg equation. Six haplotypes were determined for three PXR SNPs, and four for CAR SNPs. Tests for linkage disequilibrium showed a high association between PXR (rs38114057) and PXR (rs3814058) (p= 0.001), and between the three CAR SNPs (p=0.001), which could be useful for identification of tag SNPs.

Conclusions:In the present study, the genotype frequencies of some SNPs related to mirtazapine metabolism in Spaniards were analyzed and showed that our study population is representative of HapMap European population. The results obtained could be analyzed with pharmacokinetic parameters of mirtazapine to elucidate the genotype-phenotype relationship, the involvement of these SNPs in metabolic reactions, drug interactions, and prediction of treatment response.

Clin Chem Lab Med 2010;48:635–9.
Development of a low-cost real-time reverse transcriptase-polymerase chain reaction technique for the detection and quantification of hepatitis C viral load
Authors: Shahzamani Kiana , Merat Shahin , Rezvan Houri , Mirab-Samiee Siamak , Khademi Hooman , Malekzadeh Reza , Sabahi Farzaneh
Published: March 2010

Background: It is necessary to develop a highly specific and sensitive assay to quantify the exact amount of hepatitis C virus (HCV) RNA in blood of patients with hepatitis C. For this reason, a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay for quantification of HCV RNA in human plasma was developed.

Methods: A pair of primers as well as hybridization probes were selected. A real-time RT-PCR was set up and optimized. To establish the sensitivity of the assay, a serial dilution of HCV standards and reference sera, including the six major HCV genotypes, was used. The performance of the assay was evaluated with 191 known HCV-RNA positive and 100 negative samples.

Results: The real-time assay had a sensitivity of 50 IU/mL, with a dynamic range of detection between 103 and 106 IU/mL. The coefficients of variation of threshold cycle values in intra- and inter-day-runs were <1.77% and 3.40%, respectively. Measurement of HCV-RNA positive samples yielded reproducible data with 100% specificity.

Conclusions: The high sensitivity, simplicity, reproducibility, wide dynamic range, and low cost of this real-time HCV RNA quantification makes this method especially suitable for monitoring viral load during therapy and tailoring of treatment schedules accordingly.

Clin Chem Lab Med 2010;48:777–84.