bluuebunny/crossref_metadata_2025 huggingface dataset:
Term amniotic membrane is a high throughput source for multipotent mesenchymal stem cells with the ability to differentiate into endothelial cells in vitro
Authors: Alviano Francesco , Fossati Valentina , Marchionni Cosetta , Arpinati Mario , Bonsi Laura , Franchina Michele , Lanzoni Giacomo , Cantoni Silvia , Cavallini Claudia , Bianchi Francesca , Tazzari Pier Luigi , Pasquinelli Gianandrea , Foroni Laura , Ventura Carlo , Grossi Alberto , Bagnara Gian Paolo
Published: February 2007

Background

Term Amniotic membrane (AM) is a very attractive source of Mesenchymal Stem Cells (MSCs) due to the fact that this fetal tissue is usually discarded without ethical conflicts, leading to high efficiency in MSC recovery with no intrusive procedures. Here we confirmed that term AM, as previously reported in the literature, is an abundant source of hMSCs; in particular we further investigated the AM differentiation potential by assessing whether these cells may also be committed to the angiogenic fate. In agreement with the recommendation of the International Society for Cellular Therapy, the mesenchymal cells herein investigated were named Amniotic Membrane-human Mesenchymal Stromal Cells (AM-hMSC).

Results

The recovery of hMSCs and their in vitro expansion potential were greater in amniotic membrane than in bone marrow stroma. At flow cytometry analysis AM-hMSCs showed an immunophenotypical profile, i.e., positive for CD105, CD73, CD29, CD44, CD166 and negative for CD14, CD34, CD45, consistent with that reported for bone marrow-derived MSCs. In addition, amniotic membrane-isolated cells underwent in vitro osteogenic (von Kossa stain), adipogenic (Oil Red-O stain), chondrogenic (collagen type II immunohistochemichal detection) and myogenic (RT-PCR MyoD and Myogenin expression as well as desmin immunohistochemical detection) differentiation. In angiogenic experiments, a spontaneous differentiation into endothelial cells was detected by in vitro matrigel assay and this behaviour has been enhanced through Vascular Endothelial Growth Factor (VEGF) induction. According to these findings, VEGF receptor 1 and 2 (FLT-1 and KDR) were basally expressed in AM-hMSCs and the expression of endothelial-specific markers like FLT-1 KDR, ICAM-1 increased after exposure to VEGF together with the occurrence of CD34 and von Willebrand Factor positive cells.

Conclusion

The current study suggests that AM-hMSCs may emerge as a remarkable tool for the cell therapy of multiple diseased tissues. AM-hMSCs may potentially assist both bone and cartilage repair, nevertheless, due to their angiogenic potential, they may also pave the way for novel approaches in the development of tissue-engineered vascular grafts which are useful when vascularization of ischemic tissues is required.
The small FNR regulon of Neisseria gonorrhoeae: comparison with the larger Escherichia coli FNR regulon and interaction with the NarQ-NarP regulon
Authors: Whitehead Rebekah N , Overton Tim W , Snyder Lori AS , McGowan Simon J , Smith Harry , Cole Jeff A , Saunders Nigel J
Published: February 2007

Background

Neisseria gonorrhoeae can survive during oxygen starvation by reducing nitrite to nitrous oxide catalysed by the nitrite and nitric oxide reductases, AniA and NorB. The oxygen-sensing transcription factor, FNR, is essential for transcription activation at the aniA promoter, and full activation also requires the two-component regulatory system, NarQ-NarP, and the presence of nitrite. The only other gene known to be activated by the gonococcal FNR is ccp encoding a cytochrome c peroxidase, and no FNR-repressed genes have been reported in the gonococcus. In contrast, FNR acts as both an activator and repressor involved in the control of more than 100 operons in E. coli regulating major changes in the adaptation from aerobic to anaerobic conditions. In this study we have performed a microarray-led investigation of the FNR-mediated responses in N. gonorrhoeae to determine the physiological similarities and differences in the role of FNR in cellular regulation in this species.

Results

Microarray experiments show that N. gonorrhoeae FNR controls a much smaller regulon than its E. coli counterpart; it activates transcription of aniA and thirteen other genes, and represses transcription of six genes that include dnrN and norB. Having previously shown that a single amino acid substitution is sufficient to enable the gonococcal FNR to complement an E. coli fnr mutation, we investigated whether the gonococcal NarQ-NarP can substitute for E. coli NarX-NarL or NarQ-NarP. A plasmid expressing gonococcal narQ-narP was unable to complement E. coli narQP or narXL mutants, and was insensitive to nitrate or nitrite. Mutations that progressively changed the periplasmic nitrate sensing region, the P box, of E. coli NarQ to the sequence of the corresponding region of gonococcal NarQ resulted in loss of transcription activation in response to the availability of either nitrate or nitrite. However, the previously reported ligand-insensitive ability of gonococcal NarQ, the "locked on" phenotype, to activate either E. coli NarL or NarP was confirmed.

Conclusion

Despite the sequence similarities between transcription activators of E. coli and N. gonorrhoeae, these results emphasise the fundamental differences in transcription regulation between these two types of pathogenic bacteria.
Microarray-based analysis of fish egg quality after natural or controlled ovulation
Authors: Bonnet Emilie , Fostier Alexis , Bobe Julien
Published: February 2007

Background

The preservation of fish egg quality after ovulation-control protocols is a major issue for the development of specific biotechnological processes (e.g. nuclear transfer). Depending on the species, it is often necessary to control the timing of ovulation or induce the ovulatory process. The hormonal or photoperiodic control of ovulation can induce specific egg quality defects that have been thoroughly studied. In contrast, the impact on the egg transcriptome as a result of these manipulations has received far less attention. Furthermore, the relationship between the mRNA abundance of maternally-inherited mRNAs and the developmental potential of the egg has never benefited from genome-wide studies. Thus, the present study aimed at studying the rainbow trout (Oncorhynchus mykiss) egg transcriptome after natural or controlled ovulation using 9152-cDNA microarrays.

Results

The analysis of egg transcriptome after natural or controlled ovulation led to the identification of 26 genes. The expression patterns of 17 of those genes were monitored by real-time PCR. We observed that the control of ovulation by both hormonal induction and photoperiod manipulation induced significant changes in the egg mRNA abundance of specific genes. A dramatic increase of Apolipoprotein C1 (APOC1) and tyrosine protein kinase HCK was observed in the eggs when a hormonal induction of ovulation was performed. In addition, both microarray and real-time PCR analyses showed that prohibitin 2 (PHB2) egg mRNA abundance was negatively correlated with developmental success.

Conclusion

First, we showed, for the first time in fish, that the control of ovulation using either a hormonal induction or a manipulated photoperiod can induce differences in the egg mRNA abundance of specific genes. While the impact of these modifications on subsequent embryonic development is unknown, our observations clearly show that the egg transcriptome is affected by an artificial induction of ovulation.

Second, we showed that the egg mRNA abundance of prohibitin 2 was reflective of the developmental potential of the egg.

Finally, the identity and ontology of identified genes provided significant hints that could result in a better understanding of the mechanisms associated with each type of ovulation control (i.e. hormonal, photoperiodic), and in the identification of conserved mechanisms triggering the loss of egg developmental potential.
Correlates of depressive symptoms among Latino and Non-Latino White adolescents: Findings from the 2003 California Health Interview Survey
Authors: Mikolajczyk Rafael T , Bredehorst Maren , Khelaifat Nadia , Maier Claudia , Maxwell Annette E
Published: February 2007

Background

The prevalence of depression is increasing not only among adults, but also among adolescents. Several risk factors for depression in youth have been identified, including female gender, increasing age, lower socio-economic status, and Latino ethnic background. The literature is divided regarding the role of acculturation as risk factor among Latino youth. We analyzed the correlates of depressive symptoms among Latino and Non-Latino White adolescents residing in California with a special focus on acculturation.

Methods

We performed an analysis of the adolescent sample of the 2003 California Health Interview Survey, which included 3,196 telephone-interviews with Latino and Non-Latino White adolescents between the ages of 12 and 17. Depressive symptomatology was measured with a reduced version of the Center for Epidemiologic Studies Depression Scale. Acculturation was measured by a score based on language in which the interview was conducted, language(s) spoken at home, place of birth, number of years lived in the United States, and citizenship status of the adolescent and both of his/her parents, using canonical principal component analysis. Other variables used in the analysis were: support provided by adults at school and at home, age of the adolescent, gender, socio-economic status, and household type (two parent or one parent household).

Results

Unadjusted analysis suggested that the risk of depressive symptoms was twice as high among Latinos as compared to Non-Latino Whites (10.5% versus 5.5 %, p < 0.001). The risk was slightly higher in the low acculturation group than in the high acculturation group (13.1% versus 9.7%, p = 0.12). Similarly, low acculturation was associated with an increased risk of depressive symptoms in multivariate analysis within the Latino subsample (OR 1.54, CI 0.97–2.44, p = 0.07). Latino ethnicity emerged as risk factor for depressive symptoms among the strata with higher income and high support at home and at school. In the disadvantaged subgroups (higher poverty, low support at home and at school) Non-Latino Whites and Latinos had a similar risk of depressive symptoms.

Conclusion

Our findings suggest that the differences in depressive symptoms between Non-Latino Whites and Latino adolescents disappear at least in some strata after adjusting for socio-demographic and social support variables.
Progress towards early detection services for infants with hearing loss in developing countries
Authors: Olusanya Bolajoko O , Swanepoel De Wet , Chapchap Mônica J , Castillo Salvador , Habib Hamed , Mukari Siti Z , Martinez Norberto V , Lin Hung-Ching , McPherson Bradley
Published: February 2007

Background

Early detection of infants with permanent hearing loss through infant hearing screening is recognised and routinely offered as a vital component of early childhood care in developed countries. This article investigates the initiatives and progress towards early detection of infants with hearing loss in developing countries against the backdrop of the dearth of epidemiological data from this region.

Methods

A cross-sectional, descriptive study based on responses to a structured questionnaire eliciting information on the nature and scope of early hearing detection services; strategies for financing services; parental and professional attitudes towards screening; and the performance of screening programmes. Responses were complemented with relevant data from the internet and PubMed/Medline.

Results

Pilot projects using objective screening tests are on-going in a growing number of countries. Screening services are provided at public/private hospitals and/or community health centres and at no charge only in a few countries. Attitudes amongst parents and health care workers are typically positive towards such programmes. Screening efficiency, as measured by referral rate at discharge, was generally found to be lower than desired but several programmes achieved other international benchmarks. Coverage is generally above 90% but poor follow-up rates remain a challenge in some countries. The mean age of diagnosis is usually less than six months, even for community-based programmes.

Conclusion

Lack of adequate resources by many governments may limit rapid nationwide introduction of services for early hearing detection and intervention, but may not deter such services altogether. Parents may be required to pay for services in some settings in line with the existing practice where healthcare services are predominantly financed by out-of-pocket spending rather than public funding. However, governments and their international development partners need to complement current voluntary initiatives through systematic scaling-up of public awareness and requisite manpower development towards sustainable service capacities at all levels of healthcare delivery.
Gene expression profiling of cutaneous wound healing
Authors: Deonarine Kavita , Panelli Monica C , Stashower Mitchell E , Jin Ping , Smith Kina , Slade Herbert B , Norwood Christopher , Wang Ena , Marincola Francesco M , Stroncek David F
Published: February 2007

Background

Although the sequence of events leading to wound repair has been described at the cellular and, to a limited extent, at the protein level this process has yet to be fully elucidated. Genome wide transcriptional analysis tools promise to further define the global picture of this complex progression of events.

Study Design

This study was part of a placebo-controlled double-blind clinical trial in which basal cell carcinomas were treated topically with an immunomodifier – toll-like receptor 7 agonist: imiquimod. The fourteen patients with basal cell carcinoma in the placebo arm of the trial received placebo treatment consisting solely of vehicle cream. A skin punch biopsy was obtained immediately before treatment and at the end of the placebo treatment (after 2, 4 or 8 days). 17.5K cDNA microarrays were utilized to profile the biopsy material.

Results

Four gene signatures whose expression changed relative to baseline (before wound induction by the pre-treatment biopsy) were identified. The largest group was comprised predominantly of inflammatory genes whose expression was increased throughout the study. Two additional signatures were observed which included preferentially pro-inflammatory genes in the early post-treatment biopsies (2 days after pre-treatment biopsies) and repair and angiogenesis genes in the later (4 to 8 days) biopsies. The fourth and smallest set of genes was down-regulated throughout the study. Early in wound healing the expression of markers of both M1 and M2 macrophages were increased, but later M2 markers predominated.

Conclusion

The initial response to a cutaneous wound induces powerful transcriptional activation of pro-inflammatory stimuli which may alert the host defense. Subsequently and in the absence of infection, inflammation subsides and it is replaced by angiogenesis and remodeling. Understanding this transition which may be driven by a change from a mixed macrophage population to predominately M2 macrophages, may help the interpretation of the cellular and molecular events occurring in the microenvironment of serially biopsied tissues.
Retrospective analysis of protein kinase C-beta (PKC-β) expression in lymphoid malignancies and its association with survival in diffuse large B-cell lymphomas
Authors: Li Shuyu , Phong Mark , Lahn Michael , Brail Leslie , Sutton Susan , Lin Boris K , Thornton Donald , Liao Birong
Published: February 2007

Background

Both mechanistic features and recent correlative findings suggest a potential role for protein kinase C-beta (PKC-β) in tumor pathogenesis, particularly in B-cell malignancies. To evaluate the role of this gene in lymphoid malignancies, we analyzed global gene expression data to quantify PKC-β expression across diagnostic groups and, when possible, determined correlations between PKC-β expression and survival.

Results

Our analysis showed that the level of PKC-β expression was highest in chronic lymphocytic leukemia and follicular lymphoma. Within diffuse large-B cell lymphoma (DLBCL), PKC-β expression was significantly higher in activated B-cell- like subtype than germinal center B-cell- like subtype (P< 0.0001). Elevated PKC-β appeared to be associated with worse survival in both of these subtypes. When analyzed within clinically defined risk groups established by the International Prognostic Index (IPI), PKC-β expression was lowest in patients with low IPI scores (0–1). Within intermediate- and high-risk IPI groups, elevated PKC-β expression was associated with worse survival, suggesting that PKC-β may expand the prognostic value of the IPI. Results of global gene expression analyses of DLBCL samples corroborate previous observations that anti-apoptosis, cell proliferation, and B-cell proliferation signaling pathways are functionally related to PKC-β.

Conclusion

We present a first detailed pharmacogenomics report comparing PKC-β mRNA expression across different lymphoid malignancies and evaluating it as an outcome predictor. Our findings suggest that DLBCL patients with elevated PKC-β have a worse prognosis, indicating that further evaluation of PKC-β as a chemotherapeutic target for lymphoid malignancies is warranted.

Reviewers

This article was reviewed by Dr. Pierre Pontarotti, Dr. Kateryna Makova, and Dr. Matthew Coleman (nominated by Dr. Sandrine Dudoit).
Reliability and validity of a Dutch version of the Leicester Cough Questionnaire
Authors: Huisman Arnold N , Wu Mei-Zei , Uil Steven M , van den Berg Jan Willem K
Published: February 2007

Background

Chronic cough is a common condition with a significant impact on quality of life. Currently, no health status measure specific for chronic cough exists in the Netherlands. Thus we developed a Dutch version of the Leicester Cough Questionnaire (LCQ) and tested its scaling and clinical properties.

Methods

The LCQ was adapted for Dutch conditions following a forward-backward translation procedure. All patients referred to our cough clinic between May 2004 and February 2005 completed five questionnaires, the LCQ, the modified Borg score for cough, the Short-Form 36 (SF-36), the Hospital Anxiety and Depression Scale (HADS) and the Global Rating of Change (GRC) upon presentation, after two weeks and after 6 months. Concurrent validation, internal consistency, repeatability and responsiveness were determined.

Results

For the concurrent validation the correlation coefficients (n = 152 patients) between the LCQ and the other outcome measures varied between 0.22 and 0.61. The internal consistency of the LCQ (n = 58) was high for each of the domains with a Crohnbach's alpha coefficient between 0.77 and 0.91. The two week repeatability of the LCQ in patients with no change in cough (n = 48) was high with intraclass correlation coefficients varying between 0.86 and 0.93. Patients who reported an improvement in cough (n = 140) after 6 months demonstrated significant improvement on each of the domains of the LCQ.

Conclusion

The Dutch version of the LCQ is a valid and reliable questionnaire to measure (changes of) health status in patients with chronic cough.
Managing burn victims of suicide bombing attacks: outcomes, lessons learnt, and changes made from three attacks in Indonesia
Authors: Chim Harvey , Yew Woon Si , Song Colin
Published: February 2007

Introduction

Terror attacks in Southeast Asia were almost nonexistent until the 2002 Bali bomb blast, considered the deadliest attack in Indonesian history. Further attacks in 2003 (Jakarta), 2004 (Jakarta), and 2005 (Bali) have turned terrorist attacks into an ever-present reality.

Methods

The authors reviewed medical charts of victims evacuated to the Singapore General Hospital (SGH) Burns Centre during three suicide attacks involving Bali (2002 and 2005) and the Jakarta Marriott hotel (2003). Problems faced, lessons learnt, and costs incurred are discussed. A burns disaster plan drawing on lessons learnt from these attacks is presented.

Results

Thirty-one patients were treated at the SGH Burns Centre in three attacks (2002 Bali attack [n = 15], 2003 Jakarta attack [n = 14], and 2005 Bali attack [n = 2]). For the 2002 Bali attack, median age was 29 years (range 20 to 50 years), median percentage of total burn surface area (TBSA) was 29% (range 5% to 55%), and median abbreviated burn severity index (ABSI) was 6 (range 3 to 10). Eight of 15 patients were admitted to the intensive care unit. For the 2003 Jakarta attack, median age was 35 years (range 24 to 56 years), median percentage of TBSA was 10% (range 2% to 46%), and median ABSI was 4 (range 3 to 9). A large number of patients had other injuries. Problems faced included manpower issues, lack of bed space, shortage of blood products, and lack of cadaver skin.

Conclusion

The changing nature of terror attacks mandates continued vigilance and disaster preparedness. The multidimensional burns patient, complicated by other injuries, is likely to become increasingly common. A burns disaster plan with emphasis on effective command, control, and communication as well as organisation of health care personnel following a 'team concept' will do much to ensure that the sudden onset of a crisis situation at an unexpected time does not overwhelm hospital manpower and resources.
Revised Pediatric Emergency Assessment Tool (RePEAT): A Severity Index for Pediatric Emergency Care
Authors: Gorelick Marc H. , Alessandrini Evaline A. , Cronan Kathleen , Shults Justine
Published: February 2007

Objectives:
To develop and validate a multivariable model, using information available at the time of patient triage, to predict the level of care provided to pediatric emergency patients for use as a severity of illness measure.

Methods:
This was a retrospective cohort study of 5,521 children 18 years of age or younger treated at four emergency departments (EDs) over a 12‐month period. Data were obtained from abstraction of patient records. Logistic regression was used to develop (75% of sample) and validate (25% of sample) models to predict any nonroutine diagnostic or therapeutic intervention in the ED and admission to the hospital. Data on ED length of stay and hospital costs were also obtained.

Results:
Eight predictor variables were included in the final models: presenting complaint, age, triage acuity category, arrival by emergency medical services, current use of prescription medications, and three triage vital signs (heart rate, respiratory rate, and temperature). The resulting models had adequate goodness of fit in both derivation and validation samples. The area under the receiver operating characteristic curve was 0.73 for the ED intervention model and 0.85 for the admission model. The Revised Pediatric Emergency Assessment Tool (RePEAT) score was then calculated as the sum of the predicted probability of receiving intervention and twice the predicted probability of admission. The RePEAT score had a significant univariate association with ED costs (r= 0.44) and with ED length of stay (r= 0.27) and contributed significantly to the fit of multivariable models comparing these outcomes across sites.

Conclusions:
The RePEAT score accurately predicts level of care provided for pediatric emergency patients and may provide a useful means of risk adjustment when benchmarking outcomes.
Postmenopausal Hormone Therapy and Changes in Mammographic Density
Authors: van Duijnhoven Fränzel J.B. , Peeters Petra H.M. , Warren Ruth M.L. , Bingham Sheila A. , van Noord Paulus A.H. , Monninkhof Evelyn M. , Grobbee Diederick E. , van Gils Carla H.
Published: February 2007

Purpose

Hormone therapy (HT) use has been associated with an increased breast cancer risk. We explored the underlying mechanism further by determining the effects of HT on mammographic density, a measure of dense tissue in the breast and a consistent breast cancer risk factor.

Patients and Methods

A total of 620 HT users and 620 never users from the Dutch Prospect–European Prospective Investigation into Cancer and Nutrition (EPIC) cohort and 175 HT users and 161 never users from the United Kingdom EPIC–Norfolk cohort were included. For HT users, one mammogram before and one mammogram during HT use was included. For never users, mammograms with similar time intervals were included. Mammographic density was assessed using a computer-assisted method. Changes in density were analyzed using linear regression.

Results

The median time between mammograms was 3.0 years and the median duration of HT use was 1 year. The absolute mean decline in percent density was larger in never users (7.3%) than in estrogen therapy users (6.4%; P = .22) and combined HT users (3.5%; P < .01). The effect of HT appeared to be high in a small number of women, whereas most women were unaffected.

Conclusion

Our results suggest that HT use, and especially estrogen and progestin use, slows the changes from dense patterns to more fatty patterns that are normally seen in women with increasing age. Given that it is postulated that lifetime cumulative exposure to high density may be related to breast cancer risk, a delay in density decline in HT users potentially could explain their increased breast cancer risk.
Randomized, Double-Blind, Placebo-Controlled Trial of Erythropoietin in Non–Small-Cell Lung Cancer With Disease-Related Anemia
Authors: Wright James R. , Ung Yee C. , Julian Jim A. , Pritchard Kathleen I. , Whelan Timothy J. , Smith Column , Szechtman Barbara , Roa Wilson , Mulroy Liam , Rudinskas Leona , Gagnon Bruno , Okawara Gord S. , Levine Mark N.
Published: February 2007

Purpose

Previous trials have suggested a quality-of-life (QOL) improvement for anemic cancer patients treated with erythropoietin, but few used QOL as the primary outcome. We designed a trial to investigate the effects of epoetin alfa therapy on the QOL of anemic patients with advanced non–small-cell carcinoma of the lung (NSCLC).

Patients and Methods

A multicenter, randomized, double-blind, placebo-controlled trial was conducted. The proposed sample size was 300 patients. Eligible patients were required to have NSCLC unsuitable for curative therapy and baseline hemoglobin (Hgb) levels less than 121 g/L. Patients were assigned to 12 weekly injections of subcutaneous epoetin alpha or placebo, targeting Hgb levels between 120 and 140 g/L. The primary outcome was the difference in the change in Functional Assessment of Cancer Therapy–Anemia scores between baseline and 12 weeks.

Results

Reports of thrombotic events in other epoetin trials prompted an unplanned safety analysis after 70 patients had been randomly assigned (33 to the active arm and 37 to the placebo arm). This revealed a significant difference in the median survival in favor of the patients on the placebo arm of the trial (63 v 129 days; hazard ratio, 1.84; P = .04). The Steering Committee closed the trial. Patient numbers compromised the interpretation of the QOL analysis, but a positive Hgb response was noted with epoetin alfa treatment.

Conclusion

An unplanned safety analysis suggested decreased overall survival in patients with advanced NSCLC treated with epoetin alfa. Although infrequent, other similar reports highlight the need for ongoing trials evaluating erythropoietin receptor agonists to ensure that overall survival is monitored closely.
High-Dose [131I]Tositumomab (anti-CD20) Radioimmunotherapy and Autologous Hematopoietic Stem-Cell Transplantation for Adults ≥ 60 Years Old With Relapsed or Refractory B-Cell Lymphoma
Authors: Gopal Ajay K. , Rajendran Joseph G. , Gooley Ted A. , Pagel John M. , Fisher Darrell R. , Petersdorf Stephen H. , Maloney David G. , Eary Janet F. , Appelbaum Frederick R. , Press Oliver W.
Published: February 2007

Purpose

The majority of patients with relapsed or refractory B-cell non-Hodgkin's lymphoma (NHL) are older than 60 years, yet they are often denied potentially curative high-dose therapy and autologous stem-cell transplantations (ASCT) because of the risk of excessive treatment-related morbidity and mortality. Myeloablative anti-CD20 radioimmunotherapy (RIT) can deliver curative radiation doses to tumor sites while limiting exposure to normal organs and may be particularly suited for older adults requiring high-dose therapy.

Patients and Methods

Patients older than 60 years with relapsed B-cell NHL (B-NHL) received infusions of tositumomab anti-CD20 antibody labeled with 185 to 370 Mbq (5 to 10 mCi) [131I]-tracer for dosimetry purposes followed 10 days later by individualized therapeutic infusions of [131I]tositumomab (median, 19.4 Gbq [525 mCi]; range, 12.1 to 42.7 Gbq [328 to 1,154 mCi]) to deliver 25 to 27 Gy to the critical normal organ receiving the highest radiation dose. ASCT was performed approximately 2 weeks after therapy.

Results

Twenty-four patients with a median age of 64 years (range, 60 to 76 years), who had received a median of four prior regimens (range, two to 14 regimens), were treated. Thirteen patients (54%) had chemotherapy-resistant disease. The estimated 3-year overall and progression-free survival rates were 59% and 51%, respectively, with a median follow-up of 2.9 years (range, 1 to 6 years). All patients experienced expected myeloablation with engraftment of platelets (≥ 20 K/μL) and neutrophils (≥ 500/μL), occurring at a median of 9 and 15 days after ASCT, respectively. There were no treatment-related deaths, and only two patients experienced grade 4 nonhematologic toxicity.

Conclusion

Myeloablative RIT and ASCT is a safe and effective therapeutic option for older adults with relapsed B-NHL.
Differential Cell-Specific Modulation of HOXA10 by Estrogen and Specificity Protein 1 Response Elements
Authors: Martin Ryan , Taylor Melissa B. , Krikun Graciela , Lockwood Charles , Akbas G. Edda , Taylor Hugh S.
Published: February 2007

Context: HOX genes are highly evolutionarily conserved regulators of embryonic development. HOXA10 also regulates differentiation of the adult reproductive tract and mammary gland in response to sex steroids.

Objective: We recently identified two HOXA10 estrogen response elements (EREs). Here we demonstrate that estrogen-responsive HOXA10 expression is cell type specific.

Design and Setting: We conducted an in vitro study at an academic medical center.

Main Outcome Measure: Reporter assay, gel shift assays (electrophoretic mobility shift assay), and immunohistochemistry were done.

Results: The HOXA10 EREs and a specificity protein 1 (Sp1) binding site differentially drive the cell-type-specific E2 response. In electrophoretic mobility shift assays, both estrogen receptor-α and -β bound both EREs but not the Sp1 site. In reporter assays, both EREs and the Sp1 site demonstrated estrogen responsiveness and tissue specificity; transiently transfected uterine Ishikawa cells or breast MCF-7 cells showed differential responses to E2 treatment. Each response element (Sp1, ERE1, and ERE2) drove distinct differential expression in each cell type. Sp1 protein was expressed in a menstrual-cycle stage-specific expression pattern in endometrium, first expressed in perivascular cells.

Conclusions: Tissue specificity inherent to a regulatory element as well as differential cellular expression of transcription factors imparts differential tissue-specific estrogen responsiveness.
Novel GALNT3 Mutations Causing Hyperostosis-Hyperphosphatemia Syndrome Result in Low Intact Fibroblast Growth Factor 23 Concentrations
Authors: Ichikawa Shoji , Guigonis Vincent , Imel Erik A. , Courouble Mélanie , Heissat Sophie , Henley John D. , Sorenson Andrea H. , Petit Barbara , Lienhardt Anne , Econs Michael J.
Published: February 2007

Context: Hyperostosis-hyperphosphatemia syndrome (HHS) is a rare metabolic disorder characterized by hyperphosphatemia and localized hyperostosis. HHS is caused by mutations in GALNT3, which encodes UDP-N-acetyl-α-d-galactosamine:polypeptide N- acetylgalactosaminyltransferase 3. Familial tumoral calcinosis (TC), characterized by ectopic calcifications and hyperphosphatemia, is caused by mutations in the GALNT3 or fibroblast growth factor 23 (FGF23) genes.

Objective: Our objective was to identify mutations in FGF23 or GALNT3 and determine serum FGF23 levels in an HHS patient.

Design: Mutation detection in FGF23 and GALNT3 was performed by DNA sequencing, and serum FGF23 concentrations were measured by ELISA.

Patients or Other Participants: A 5-year-old French boy with HHS and his family members participated.

Results: The patient presented with painful cortical lesions in his leg. Radiographs of the affected bone showed diaphyseal hyperostosis. The lesional tissue comprised trabeculae of immature, woven bone surrounded by fibrous tissue. Biochemistry revealed elevated phosphate, tubular maximum rate for phosphate reabsorption per deciliter of glomerular filtrate, and 1,25-dihydroxyvitamin D levels. The patient was a compound heterozygote for two novel GALNT3 mutations. His parents and brother were heterozygous for one of the mutations and had no biochemical abnormalities. Intact FGF23 level in the patient was low normal, whereas C-terminal FGF23 was elevated, a pattern similar to TC.

Conclusion: The presence of GALNT3 mutations and elevated C-terminal, but low intact serum FGF23, levels in HHS resemble those seen in TC, suggesting that HHS and TC are different manifestations of the same disorder. The absence of biochemical abnormalities in the heterozygous individuals suggests that one normal allele is sufficient for secretion of intact FGF23.
Uncomplicated Type 1 Diabetes Is Associated with Increased Asymmetric Dimethylarginine Concentrations
Authors: Altinova Alev E. , Arslan Metin , Sepici-Dincel Aylin , Akturk Mujde , Altan Nilgun , Toruner Fusun B.
Published: February 2007

Context: Asymmetric dimethylarginine (ADMA) has recently emerged as an independent risk marker for cardiovascular disease, but studies investigating the ADMA levels in type 1 diabetes mellitus (DM) are scarce.

Objective: We aimed to evaluate plasma ADMA, l-arginine concentrations, and l-arginine to ADMA ratio in uncomplicated type 1 diabetic patients and controls.

Design and Subjects: Forty patients with type 1 DM who did not have clinical evidence of vascular complications and 35 healthy controls were included in the study.

Results: Plasma ADMA concentrations were higher (2.6 ± 1.9 vs. 1.7 ± 0.7 μmol/liter, P < 0.01), and l-arginine levels were lower (79.3 ± 22.6 vs. 89.6 ± 19.4 μmol/liter, P < 0.05) in the diabetic group, compared with controls. The l-arginine to ADMA ratio was also lower in the diabetic group (38.7 ± 17.1 vs. 62.0 ± 27.9, P < 0.0001). In diabetic patients, logADMA correlated positively with body mass index (BMI) (P = 0.01), fasting blood glucose (P = 0.006), and low-density lipoprotein cholesterol (LDL-c) (P = 0.01) and negatively with high-density lipoprotein cholesterol (P = 0.03). l-Arginine to ADMA ratio correlated negatively with BMI (P = 0.004), fasting blood glucose (P = 0.02), and LDL-c (P = 0.01) and positively with high-density lipoprotein cholesterol (P = 0.04). In controls, logADMA and l-arginine to ADMA ratio correlated with BMI and LDL-c (P < 0.05). In regression analysis, BMI predicted 15% variance of ADMA levels (P = 0.02).

Conclusions: We demonstrated that ADMA increases and l-arginine to ADMA ratio decreases, even before the development of vascular complications in type 1 DM.
Disrupted mechanical stability of the dystrophin-glycoprotein complex causes severe muscular dystrophy in sarcospan transgenic mice
Authors: Peter Angela K. , Miller Gaynor , Crosbie Rachelle H.
Published: February 2007

The dystrophin-glycoprotein complex spans the muscle plasma membrane and provides a mechanical linkage between laminin in the extracellular matrix and actin in the intracellular cytoskeleton. Within the dystrophin-glycoprotein complex, the sarcoglycans and sarcospan constitute a subcomplex of transmembrane proteins that stabilize α-dystroglycan, a receptor for laminin and other components of the extracellular matrix. In order to elucidate the function of sarcospan, we generated transgenic mice that overexpress sarcospan in skeletal muscle. Sarcospan transgenic mice with moderate (tenfold) levels of sarcospan overexpression exhibit a severe phenotype that is similar to mouse models of laminin-deficient congenital muscular dystrophy (MD). Sarcospan transgenic mice display severe kyphosis and die prematurely between 6 and 10 weeks of age. Histological analysis reveals that sarcospan expression causes muscle pathology marked by increased muscle fiber degeneration and/or regeneration. Sarcospan transgenic muscle does not display sarcolemma damage, which is distinct from dystrophin- and sarcoglycan-deficient muscular dystrophies. We show that sarcospan clusters the sarcoglycans into insoluble protein aggregates and causes destabilization of α-dystroglycan. Evidence is provided to demonstrate abnormal extracellular matrix assembly, which represents a probable pathological mechanism for the severe and lethal dystrophic phenotype. Taken together, these data suggest that sarcospan plays an important mechanical role in stabilizing the dystrophin-glycoprotein complex.
The lissencephaly protein Lis1 is present in motile mammalian cilia and requires outer arm dynein for targeting to Chlamydomonas flagella
Authors: Pedersen Lotte B. , Rompolas Panteleimon , Christensen Søren T. , Rosenbaum Joel L. , King Stephen M.
Published: February 2007

Lissencephaly is a developmental brain disorder characterized by a smooth cerebral surface, thickened cortex and misplaced neurons. Classical lissencephaly is caused by mutations in LIS1, which encodes a WD-repeat protein involved in cytoplasmic dynein regulation, mitosis and nuclear migration. Several proteins required for nuclear migration in Aspergillus bind directly to Lis1, including NudC. Mammalian NudC is highly expressed in ciliated epithelia, and localizes to motile cilia in various tissues. Moreover, a NudC ortholog is upregulated upon deflagellation in Chlamydomonas. We found that mammalian Lis1 localizes to motile cilia in trachea and oviduct, but is absent from non-motile primary cilia. Furthermore, we cloned a gene encoding a Lis1-like protein (CrLis1) from Chlamydomonas. CrLis1 is a ∼37 kDa protein that contains seven WD-repeat domains, similar to Lis1 proteins from other organisms. Immunoblotting using an anti-CrLis1 antibody revealed that this protein is present in the flagellum and is depleted from flagella of mutants with defective outer dynein arm assembly, including one strain that lacks only the α heavy chain/light chain 5 thioredoxin complex. Biochemical experiments confirmed that CrLis1 associates with outer dynein arm components and revealed that CrLis1 binds directly to rat NudC. Our results suggest that Lis1 and NudC are present in cilia and flagella and may regulate outer dynein arm activity.
Note—Computing Time-Dependent Waiting Time Probabilities in M(t)/M/s(t) Queuing Systems
Authors: Green Linda V. , Soares João
Published: February 2007

In this note we present algorithms that compute, exactly or approximately, time-dependent waiting time tail probabilities and the time-dependent expected waiting time in M(t)/M/s(t) queuing systems.
Progenitor Cell Isolation from Muscle-derived Cells based on Adhesion Properties
Authors: Rouger Karl , Fornasari Benoît , Armengol Valerie , Jouvion Gregory , Leroux Isabelle , Dubreil Laurence , Feron Marie , Guevel Laetitia , Cherel Yan
Published: February 2007

Adult skeletal muscle possesses remarkable regenerative capacity that has conventionally been attributed to the satellite cells. These precursor cells were thought to contain distinct populations with varying myogenic potential. Recently, the identification of multipotent stem cells capable of new myofiber formation has expanded the general view on the muscle regenerative process. Here we examined the characteristics of turkey skeletal muscle-derived cell (MDC) populations that were separated according to their adhesion abilities. We sought to determine whether these abilities could be a potential tool for separating cells with different myogenic commitment. Using the preplate technique, we showed that MDCs display a wide range of adhesion ability, allowing us to isolate a marginal fraction with initial adhesion defect. Methodological investigations revealed that this defect represents an intrinsic and well-established biological feature for these cells. In vitro behavioral and morphological analyses showed that late adherent cells (LACs) share several primitive cell characteristics. Phenotypic assessment indicated that LACs contain early stage myogenic cells and immature progenitors of satellite cells, whereas early adherent cells consist mainly of fully committed precursors. Overall, our findings demonstrate for the first time in an avian model that differential MDC adhesion properties could be used to efficiently purify cells with varying myogenic commitment, including immature progenitor cells. This manuscript contains online supplemental material at http://www.jhc.org . Please visit this article online to view these materials.
VGF Metabolic-related Gene
Authors: Cocco Cristina , Brancia Carla , Pirisi Ivo , D'Amato Filomena , Noli Barbara , Possenti Roberta , Ferri Gian-Luca
Published: February 2007

The vgf gene has been shown to be involved in several metabolic pathways. Because the pancreas is crucial to metabolism and food intake, we studied the VGF peptides in bovine, rat, and pig Langherans islets using antisera raised against specific sites along the primary sequence of the rat/mouse and human VGF protein precursor. Whereas almost all of the pancreatic endocrine cells expressed vgf mRNA, when using the VGF antisera a different staining pattern became apparent. VGF556–565 and VGF282–291 immunoreactivity were exclusively found in δ somatostatin-producing cells, whereas the human C-terminus antiserum selectively immunolabeled α glucagon and pancreatic polypeptide cells. The same cells were decorated with the VGF443–588 antiserum, which also weakly labeled β insulin-secreting cells. Finally, the VGF298–306 peptide and the rat C terminus were found in virtually all pancreatic endocrine cells. Using bovine, swine, and rat pancreatic extracts, data from chromatography and ELISA assay showed the presence of a high molecular mass form compatible with the proVGF and lower molecular mass fractions corresponding to short VGF peptides. In conclusion, selective VGF distribution may suggest a multifaceted cell type-specific processing of proVGF, resulting indifferent peptides probably involvedin neuroendocrine regulatory metabolic mechanisms.
A Differential Ligand-mediated Response of Green Fluorescent Protein-tagged Androgen Receptor in Living Prostate Cancer and Non-prostate Cancer Cell Lines
Authors: Nakauchi Hiroo , Matsuda Ken-ichi , Ochiai Ikuo , Kawauchi Akihiro , Mizutani Yoichi , Miki Tsuneharu , Kawata Mitsuhiro
Published: February 2007

Androgen has been shown to promote the proliferation of prostate cancer through the action of the androgen receptor (AR). Mutation (T877A) of the AR gene found in an androgen-sensitive prostate cancer cell line, LNCaP, has been postulated to be involved in hypersensitivity and loss of specificity for androgen. In the present study, trafficking of AR and AR (T877A) in living prostate and non-prostate cancer cell lines under high and low concentrations of androgen and antiandrogen was investigated by tagging green fluorescent protein (GFP) to the receptors. In the presence of a high concentration of androgen, AR-GFP localized in the nucleus by forming discrete clusters in all cell lines. AR (T877A)-GFP was also translocated to the nucleus in LNCaP and COS-1 cells by the addition of a high concentration of androgen. In contrast, in the presence of a low concentration of androgen, the translocation of AR-GFP and AR (T877A)-GFP was observed in LNCaP cells, but not in COS-1 cells. Upon the addition of antiandrogen, AR-GFP was translocated to the nucleus but did not form subnuclear foci in both COS-1 and LNCaP cells, whereas AR (T877A)-GFP in both cells was translocated to the nucleus with subnuclear foci. The present study demonstrates the differential response of nuclear trafficking of AR and its mutant in prostate cancer cell lines and COS cells, and the subcellular and subnuclear compartmentalization provide important information on the sensitivity of the AR mutation.
Evidence for p53 as Guardian of the Cardiomyocyte Mitochondrial Genome Following Acute Adriamycin Treatment
Authors: Nithipongvanitch Ramaneeya , Ittarat Wanida , Velez Joyce M. , Zhao Rui , St. Clair Daret K. , Oberley Terry D.
Published: February 2007

The present study is an initial analysis of whether p53 may function as guardian of the cardiomyocyte mitochondrial genome, with mitochondrial p53 localization proposed to be involved in both mitochondrial DNA (mtDNA) repair and apoptosis. Subcellular distribution, protein levels, and possible function(s) of p53 protein in the response of cardiomyocytes to adriamycin (ADR) were analyzed. Levels and subcellular localization of proteins were determined by Western blot and immunogold ultrastructural analysis techniques. Here we demonstrate that stress caused by ADR induced upregulation of p53 protein in cardio-myocyte mitochondria and nuclei between 3 and 24 hr. Increased expression of PUMA and Bax proteins, pro-apoptotic targets of p53, was documented following ADR treatment and was accompanied by increased levels of apoptotic markers, with elevation of cytosolic cytochrome c at 24 hr and subsequent caspase-3 cleavage at 3 days. Mitochondrial p53 levels correlated with mtDNA oxidative damage. Loss of p53 in knockout mouse heart resulted in a significant increase in mtDNA vulnerability to damage following ADR treatment. Our results suggest that mitochondrial p53 could participate in mtDNA repair as a first response to oxidative damage of cardiomyocyte mtDNA and demonstrate an increase of apoptotic markers as a result of mitochondrial/nuclear p53 localization.
Robust Immunohistochemical Staining of Several Classes of Proteins in Tissues Subjected to Autolysis
Authors: Maleszewski Joseph , Lu Jie , Fox-Talbot Karen , Halushka Marc K.
Published: February 2007

Despite the common use of immunohistochemistry in autopsy tissues, the stability of most proteins over extended time periods is unknown. The robustness of signal for 16 proteins (MMP1, MMP2, MMP3, MMP9, TIMP1, TIMP2, TIMP3, AGER, MSR, SCARB1, OLR1, CD36, LTF, LGALS3, LYZ, and DDOST) and two measures of advanced glycation end products (AGE, CML) was evaluated. Two formalin-fixed, paraffin-embedded human tissue arrays containing 16 tissues each were created to evaluate 48 hr of autolysis in a warm or cold environment. For these classes of proteins, matrix metalloproteinases and their inhibitors, scavenger receptors, and advanced glycation end product receptors, we saw no systematic diminution of signal intensity during a period of 24 hr. Analysis was performed by two independent observers and confirmed for a subset of proteins by digital analysis and Western blotting. We conclude that these classes of proteins degrade slowly and faithfully maintain their immunohistochemistry characteristics over at least a 24-hr time interval in devitalized tissues. This study supports the use of autopsy tissues with short postmortem intervals for immunohistochemical studies for diseases such as diabetic vascular disease, cancer, Alzheimer's disease, atherosclerosis, and other pathological states. This manuscript contains online supplemental material at http://www.jhc.org . Please visit this article online to view these materials.
Renal impairment compromises the use of total homocysteine and methylmalonic acid but not total vitamin B12 and holotranscobalamin in screening for vitamin B12 deficiency in the aged
Authors: Loikas Saila , Koskinen Pertti , Irjala Kerttu , Löppönen Minna , Isoaho Raimo , Kivelä Sirkka-Liisa , Pelliniemi Tarja-Terttu
Published: February 2007

Clin Chem Lab Med 2007;45:197–201.
Diagnostic value of calculated testosterone indices in the assessment of polycystic ovary syndrome
Authors: Hahn Susanne , Kuehnel Werner , Tan Susanne , Kramer Karen , Schmidt Markus , Roesler Susanne , Kimmig Rainer , Mann Klaus , Janssen Onno E.
Published: February 2007

Clin Chem Lab Med 2007;45:202–7.