bluuebunny/crossref_metadata_2025 huggingface dataset:
Smoking, breastfeeding, physical inactivity, contact with animals, and size of the family influence the risk of inflammatory bowel disease: A Slovak case–control study
Authors: Hlavaty Tibor , Toth Jozef , Koller Tomas , Krajcovicova Anna , Oravcova Stanislava , Zelinkova Zuzana , Huorka Martin
Published: February 2013

Background

The aetiology of inflammatory bowel disease (IBD) is not known but is likely to involve a combination of genetic predisposition and environmental risk factors. Smoking has been associated consistently with a higher risk of Crohn’s disease (CD), while appendectomy and smoking appear to diminish the risk of ulcerative colitis (UC). The roles of other environmental factors are unclear. The aim of the present study was to evaluate the association of CD and UC with several environmental risk factors.

Methods

This case–control study included 338 patients (190 CD, 148 UC) and 355 controls. All subjects completed a detailed questionnaire regarding breastfeeding duration, history of helminthic infections, allergic diseases, appendectomy, household size, housing type, contact with specific domestic animals, physical activity, and smoking. Associations between risk factors and CD and UC were investigated by univariate and multivariate analysis.

Results

On multivariate analysis, CD associated with smoking at diagnosis (odds ratio, OR, 3.7, 95% CI 2.2–6.2; p  < 0.001), being breastfed for <6 months (OR 2.7, 95% CI 1.7–4.4; p  < 0.001), and less than two childhood sporting activities weekly (OR 2.7, 95% CI 1.5–5.0; p  < 0.001) and inversely associated with frequent contact with cats in childhood (OR 0.6, 95% CI 0.4–0.9; p  < 0.03). UC associated with less than two sporting weekly activities in childhood (OR 2.0, 95% CI 1.1–3.5, p  = 0.02), fewer household members in childhood (OR 0.8, 95% CI 0.7–0.98, p  = 0.03), and being breastfed for <6 months (OR 1.7, 95% CI 1.02–2.8, p  = 0.04). A composite environmental risk index for CD revealed that 47 and 14% of the controls and patients with CD had no risk factors, respectively, and that 14 and 38% of the controls and patients with CD had at least two risk factors, respectively.

Conclusion

CD and UC associated with infrequent childhood sports activities and short breastfeeding. Furthermore, CD associated with smoking and infrequent contact with animals in childhood. UC associated with a smaller family size in childhood.
The Roles of Combat Exposure, Personal Vulnerability, and Involvement in Harm to Civilians or Prisoners in Vietnam-War-Related Posttraumatic Stress Disorder
Authors: Dohrenwend Bruce P. , Yager Thomas J. , Wall Melanie M. , Adams Ben G.
Published: February 2013

The diagnosis posttraumatic stress disorder was introduced in 1980 amid debate about the psychiatric toll of the Vietnam War. There is controversy, however, about its central assumption that potentially traumatic stressors are more important than personal vulnerability in causing the disorder. We tested this assumption with data from a rigorously diagnosed male subsample ( n = 260) from the National Vietnam Veterans Readjustment Study. Combat exposure, prewar vulnerability, and involvement in harming civilians or prisoners were examined, with only combat exposure proving necessary for disorder onset. Although none of the three factors proved sufficient, estimated onset reached 97% for veterans high on all three, with harm to civilians or prisoners showing the largest independent contribution. Severity of combat exposure proved more important than prewar vulnerability in onset; prewar vulnerability was at least as important in long-term persistence. Implications for the primacy of the stressor assumption, further research, and policy are discussed.
Expressive Writing Can Impede Emotional Recovery Following Marital Separation
Authors: Sbarra David A. , Boals Adriel , Mason Ashley E. , Larson Grace M. , Mehl Matthias R.
Published: February 2013

Marital separation and divorce are common life events that increase risk for poor health outcomes, yet few intervention studies explore how to mitigate this increased risk. This study implemented an expressive writing (EW) intervention for adults who experienced a recent marital separation. Ninety participants (32 men) were randomly assigned to and completed one of three experimental writing tasks: traditional EW, a novel (narrative-based) type of EW, or control writing. Up to 9 months after this writing, participants who were judged to be actively engaged in a search for meaning concerning their separation reported significantly worse emotional outcomes when assigned to either EW condition relative to control writing. Within the control condition, those participants who were actively engaged in a search for meaning reported the lowest levels of separation-related disturbance. We discuss these results in terms of the factors that may limit and promote psychological recovery following marital separation.
A Comparison of Two Models of Urgency
Authors: Smith Gregory T. , Guller Leila , Zapolski Tamika C. B.
Published: February 2013

The aim of this study was to test two competing theories concerning the trait of urgency: (a) Urgency reflects the tendency to act rashly/impulsively when emotional, or (b) urgency reflects a general reflexive responsivity to emotions that can lead either to rash action or to ill-advised inaction and thus to either impulsive behavior or depression. Following prior findings that fifth-grade urgency predicted sixth-grade impulsive behavior, we tested whether urgency also predicted sixth-grade depression ( n = 1,906). After controlling for sex, early pubertal onset, fifth-grade depression, fifth-grade engagement in addictive behaviors, negative affect, positive affect, and other impulsivity-related traits, fifth-grade urgency level did predict higher levels of depression at the end of sixth grade. This finding is consistent with the view that urgency can lead either to rash action or to ill-advised inaction. Urgency may be of transdiagnostic importance, contributing to both internalizing and externalizing dysfunction.
Processing, rheology and characterization of polypropylene nanocomposites and their blown films
Authors: Kashyap Manash Jyoti , Ghosh Anup K
Published: February 2013

Polypropylene nanocomposites were melt blended and analyzed for both high shear and low shear rheology. Rheological characterization carried out at high shear rates showed lower viscosity values for the nanocomposites compared to the neat polypropylene. Complex viscosity values at low shear rates have shown an increasing trend on increasing nanoclay content. Zero-shear viscosity was found to increase with nanoclay loading. Steady rate sweep test results showed shear thinning of polypropylene and nanocomposites occurring in three steps. First normal stress difference showed higher values for 5 phr nanocomposite up to intermediate shear rate and subsequently lower values. Extensional viscosity experiments were carried out at different strain rates of 0.001, 0.01, 0.1 and 1 s−1. Polypropylene nanoclay samples showed better strain-hardening behavior compared to that in neat polypropylene. Polypropylene nanocomposites blown films prepared at different nip-roll speeds were characterized for their tensile properties and film in-homogeneity.
Novel core promoter elements in the oomycete pathogen Phytophthora infestansand their influence on expression detected by genome-wide analysis
Authors: Roy Sourav , Poidevin Laetitia , Jiang Tao , Judelson Howard S
Published: February 2013

Background

The core promoter is the region flanking the transcription start site (TSS) that directs formation of the pre-initiation complex. Core promoters have been studied intensively in mammals and yeast, but not in more diverse eukaryotes. Here we investigate core promoters in oomycetes, a group within the Stramenopile kingdom that includes important plant and animal pathogens. Prior studies of a small collection of genes proposed that oomycete core promoters contain a 16 to 19 nt motif bearing an Initiator-like sequence (INR) flanked by a novel sequence named FPR, but this has not been extended to whole-genome analysis.

Results

We used expectation maximization to find over-represented motifs near TSSs of Phytophthora infestans, the potato blight pathogen. The motifs corresponded to INR, FPR, and a new element found about 25 nt downstream of the TSS called DPEP. TATA boxes were not detected. Assays of DPEP function by mutagenesis were consistent with its role as a core motif. Genome-wide searches found a well-conserved combined INR+FPR in only about 13% of genes after correcting for false discovery, which contradicted prior reports that INR and FPR are found together in most genes. INR or FPR were found alone near TSSs in 18% and 7% of genes, respectively. Promoters lacking the motifs had pyrimidine-rich regions near the TSS. The combined INR+FPR motif was linked to higher than average mRNA levels, developmentally-regulated transcription, and functions related to plant infection, while DPEP and FPR were over-represented in constitutively-expressed genes. The INR, FPR, and combined INR+FPR motifs were detected in other oomycetes including Hyaloperonospora arabidopsidis, Phytophthora sojae, Pythium ultimum, and Saprolegnia parasitica, while DPEP was found in all but S. parasitica. Only INR seemed present in a non-oomycete stramenopile.

Conclusions

The absence of a TATA box and presence of novel motifs show that the oomycete core promoter is diverged from that of model systems, and likely explains the lack of activity of non-oomycete promoters in Phytophthora transformants. The association of the INR+FPR motif with developmentally-regulated genes shows that oomycete core elements influence stage-specific transcription in addition to regulating formation of the pre-initiation complex.
Characterization of the global transcriptome for Pyropia haitanensis(Bangiales, Rhodophyta) and development of cSSR markers
Authors: Xie Chaotian , Li Bing , Xu Yan , Ji Dehua , Chen Changsheng
Published: February 2013

Background

Pyropia haitanensis is an economically important mariculture crop in China and is also valuable in life science research. However, the lack of genetic information of this organism hinders the understanding of the molecular mechanisms of specific traits. Thus, high-throughput sequencing is needed to generate a number of transcriptome sequences to be used for gene discovery and molecular marker development.

Results

In this study, high-throughput sequencing was used to analyze the global transcriptome of P. haitanensis. Approximately 103 million 90 bp paired-end reads were generated using an Illumina HiSeq 2000. De novo assembly with paired-end information yielded 24,575 unigenes with an average length of 645 bp. Based on sequence similarity searches with known proteins, a total of 16,377 (66.64%) genes were identified. Of these annotated unigenes, 5,471 and 9,168 unigenes were assigned to gene ontology and clusters of orthologous groups, respectively. Searching against the KEGG database indicated that 12,167 (49.51%) unigenes mapped to 124 KEGG pathways. Among the carbon fixation pathways, almost all the essential genes related to the C3- and C4-pathways for P. haitanensis were discovered. Significantly different expression levels of three key genes (Rubisco, PEPC and PEPCK) in different lifecycle stages of P. haitanensis indicated that the carbon fixation pathway in the conchocelis and thallus were different, and the C4-like pathway might play important roles in the conchocelis stage. In addition, 2,727 cSSRs loci were identified in the unigenes. Among them, trinucleotide SSRs were the dominant repeat motif (87.17%, 2,377) and GCC/CCG motifs were the most common repeats (60.07%, 1,638). High quality primers to 824 loci were designed and 100 primer pairs were randomly evaluated in six strains of P. haitanensis. Eighty-seven primer pairs successfully yielded amplicons.

Conclusion

This study generated a large number of putative P. haitanensis transcript sequences, which can be used for novel gene discovery and gene expression profiling analyses under different physiological conditions. A number of the cSSR markers identified can be used for molecular markers and will facilitate marker assisted selection in P. haitanensis breeding. These sequences and markers will provide valuable resources for further P. haitanensis studies.
Identification of genes involved in serum tolerance in the clinical strain Cronobacter sakazakiiES5
Authors: Schwizer Sarah , Tasara Taurai , Zurfluh Katrin , Stephan Roger , Lehner Angelika
Published: February 2013

Background

Cronobacter spp. are opportunistic pathogens that can cause septicemia and infections of the central nervous system primarily in premature, low-birth weight and/or immune-compromised neonates. Serum resistance is a crucial virulence factor for the development of systemic infections, including bacteremia. It was the aim of the current study to identify genes involved in serum tolerance in a selected Cronobacter sakazakii strain of clinical origin.

Results

Screening of 2749 random transposon knock out mutants of a C. sakazakii ES 5 library for modified serum tolerance (compared to wild type) revealed 10 mutants showing significantly increased/reduced resistance to serum killing. Identification of the affected sites in mutants displaying reduced serum resistance revealed genes encoding for surface and membrane proteins as well as regulatory elements or chaperones. By this approach, the involvement of the yet undescribed Wzy_C superfamily domain containing coding region in serum tolerance was observed and experimentally confirmed. Additionally, knock out mutants with enhanced serum tolerance were observed. Examination of respective transposon insertion loci revealed regulatory (repressor) elements, coding regions for chaperones and efflux systems as well as the coding region for the protein YbaJ. Real time expression analysis experiments revealed, that knock out of the gene for this protein negatively affects the expression of the fimA gene, which is a key structural component of the formation of fimbriae. Fimbriae are structures of high immunogenic potential and it is likely that absence/truncation of the ybaJ gene resulted in a non-fimbriated phenotype accounting for the enhanced survival of this mutant in human serum.

Conclusion

By using a transposon knock out approach we were able to identify genes involved in both increased and reduced serum tolerance in Cronobacter sakazakii ES5. This study reveals first insights in the complex nature of serum tolerance of Cronobacter spp.
Single cell transcriptomic analysis of prostate cancer cells
Authors: Welty Christopher J , Coleman Ilsa , Coleman Roger , Lakely Bryce , Xia Jing , Chen Shu , Gulati Roman , Larson Sandy R , Lange Paul H , Montgomery Bruce , Nelson Peter S , Vessella Robert L , Morrissey Colm
Published: February 2013

Background

The ability to interrogate circulating tumor cells (CTC) and disseminated tumor cells (DTC) is restricted by the small number detected and isolated (typically <10). To determine if a commercially available technology could provide a transcriptomic profile of a single prostate cancer (PCa) cell, we clonally selected and cultured a single passage of cell cycle synchronized C4-2B PCa cells. Ten sets of single, 5-, or 10-cells were isolated using a micromanipulator under direct visualization with an inverted microscope. Additionally, two groups of 10 individual DTC, each isolated from bone marrow of 2 patients with metastatic PCa were obtained. RNA was amplified using the WT-Ovation™ One-Direct Amplification System. The amplified material was hybridized on a 44K Whole Human Gene Expression Microarray. A high stringency threshold, a mean Alexa Fluor® 3 signal intensity above 300, was used for gene detection. Relative expression levels were validated for select genes using real-time PCR (RT-qPCR).

Results

Using this approach, 22,410, 20,423, and 17,009 probes were positive on the arrays from 10-cell pools, 5-cell pools, and single-cells, respectively. The sensitivity and specificity of gene detection on the single-cell analyses were 0.739 and 0.972 respectively when compared to 10-cell pools, and 0.814 and 0.979 respectively when compared to 5-cell pools, demonstrating a low false positive rate. Among 10,000 randomly selected pairs of genes, the Pearson correlation coefficient was 0.875 between the single-cell and 5-cell pools and 0.783 between the single-cell and 10-cell pools. As expected, abundant transcripts in the 5- and 10-cell samples were detected by RT-qPCR in the single-cell isolates, while lower abundance messages were not. Using the same stringency, 16,039 probes were positive on the patient single-cell arrays. Cluster analysis showed that all 10 DTC grouped together within each patient.

Conclusions

A transcriptomic profile can be reliably obtained from a single cell using commercially available technology. As expected, fewer amplified genes are detected from a single-cell sample than from pooled-cell samples, however this method can be used to reliably obtain a transcriptomic profile from DTC isolated from the bone marrow of patients with PCa.
The eCALM Trial-eTherapy for cancer appLying mindfulness: online mindfulness-based cancer recovery program for underserved individuals living with cancer in Alberta: protocol development for a randomized wait-list controlled clinical trial
Authors: Zernicke Kristin A , Campbell Tavis S , Speca Michael , McCabe-Ruff Kelley , Flowers Steven , Dirkse Dale A , Carlson Linda E
Published: February 2013

Background

Elevated stress can exacerbate cancer symptom severity, and after completion of primary cancer treatments, many individuals continue to have significant distress. Mindfulness-Based Cancer Recovery (MBCR) is an 8-week group psychosocial intervention consisting of training in mindfulness meditation and yoga designed to mitigate stress, pain, and chronic illness. Efficacy research shows face-to-face (F2F) MBCR programs have positive benefits for cancer patients; however barriers exist that impede participation in F2F groups. While online MBCR groups are available to the public, none have been evaluated. Primary objective: determine whether underserved patients are willing to participate in and complete an online MBCR program. Secondary objectives: determine whether online MBCR will mirror previous efficacy findings from F2F MBCR groups on patient-reported outcomes.

Method/design

The study includes cancer patients in Alberta, exhibiting moderate distress, who do not have access to F2F MBCR. Participants will be randomized to either online MBCR, or waiting for the next available group. An anticipated sample size of 64 participants will complete measures online pre and post treatment or waiting period. Feasibility will be tracked through monitoring numbers eligible and participating through each stage of the protocol.

Discussion

47 have completed/completing the intervention. Data suggest it is possible to conduct a randomized waitlist controlled trial of online MBCR to reach underserved cancer survivors.

Trial registration

Clinical Trials.gov Identifier: NCT01476891
Spectral broadening effects of high-power amplifiers in MIMO–OFDM relaying channels
Authors: Ahmad Ishtiaq , Sulyman Ahmed Iyanda , Alsanie Abdulhameed , Alasmari Awad Kh , Alshebeili Saleh A
Published: February 2013

The combination of MIMO–OFDM is a very attractive solution for broadband wireless services. Thus, the two prominent fourth-generation (4G) cellular systems, WiMAX and LTE-advanced, have both adopted MIMO–OFDM transmission at the physical layer. OFDM signal however suffers from nonlinear distortions when passed through high-power amplifier (HPA) at the RF stage. This nonlinear distortion introduces out-of-band spectral broadening and in-band distortions on the transmitted signals. 4G cellular standards have placed strict limits on the allowable spectral broadening in their spectrum mask specifications, to insure that data transmission on a given channel is not interfering significantly with an adjacent channel user. In this article, we characterize the out-of-band spectral broadening introduced by HPA when MIMO–OFDM signals are transmitted over multiple relaying channels. Expressions for the power spectral density of MIMO–OFDM signals are derived over multiple relay channels, and the cumulative effects of HPA on the spectrum of the transmitted signals are estimated. It is shown that depending on the number of relays and the relaying configuration employed, it may happen that a transmitted MIMO–OFDM signal with the transmit spectrum mask initially within the allowable set limit at the source node arrives at the destination violating this limit due to the cumulative effects of the multiple HPA’s in a multihop relaying channel.
Pulmonary metastasis of a papillary thyroid carcinoma and primary lung adenocarcinoma: two coincident carcinomas at the same location
Authors: Xue Liyan , Luan Zhonghua , Liu Ying , Zou Shuangmei , Jiang Jun , Wu Ning , Lu Ning , Lin Dongmei
Published: February 2013

Tumor-to-tumor metastasis is a fairly rare phenomenon. The lung cancers are the most common donors, but are exceedingly rare as recipients. Here we report a case of a lung adenocarcinoma acting as the recipient of papillary thyroid carcinoma, with multiple spreading foci of the two cancers in the lung simultaneously. The morphology and immunohistochemisty (Napsin-A, Thyroglobulin) are very important in differential diagnosis of lung primary adenocarcinoma and metastatic papillary thyroid carcinoma.

Virtual Slides

The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2069496615891134
Production of bacterial cellulose and enzyme from waste fiber sludge
Authors: Cavka Adnan , Guo Xiang , Tang Shui-Jia , Winestrand Sandra , Jönsson Leif J , Hong Feng
Published: February 2013

Background

Bacterial cellulose (BC) is a highly crystalline and mechanically stable nanopolymer, which has excellent potential as a material in many novel applications, especially if it can be produced in large amounts from an inexpensive feedstock. Waste fiber sludge, a residue with little or no value, originates from pulp mills and lignocellulosic biorefineries. A high cellulose and low lignin content contributes to making the fiber sludge suitable for bioconversion, even without a thermochemical pretreatment step. In this study, the possibility to combine production of BC and hydrolytic enzymes from fiber sludge was investigated. The BC was characterized using field-emission scanning electron microscopy and X-ray diffraction analysis, and its mechanical properties were investigated.

Results

Bacterial cellulose and enzymes were produced through sequential fermentations with the bacterium Gluconacetobacter xylinus and the filamentous fungus Trichoderma reesei. Fiber sludges from sulfate (SAFS) and sulfite (SIFS) processes were hydrolyzed enzymatically without prior thermochemical pretreatment and the resulting hydrolysates were used for BC production. The highest volumetric yields of BC from SAFS and SIFS were 11 and 10 g/L (DW), respectively. The BC yield on initial sugar in hydrolysate-based medium reached 0.3 g/g after seven days of cultivation. The tensile strength of wet BC from hydrolysate medium was about 0.04 MPa compared to about 0.03 MPa for BC from a glucose-based reference medium, while the crystallinity was slightly lower for BC from hydrolysate cultures. The spent hydrolysates were used for production of cellulase with T. reesei. The cellulase activity (CMCase activity) in spent SAFS and SIFS hydrolysates reached 5.2 U/mL (87 nkat/mL), which was similar to the activity level obtained in a reference medium containing equal amounts of reducing sugar.

Conclusions

It was shown that waste fiber sludge is a suitable raw material for production of bacterial cellulose and enzymes through sequential fermentation. The concept studied offers efficient utilization of the various components in fiber sludge hydrolysates and affords a possibility to combine production of two high value-added products using residual streams from pulp mills and biorefineries. Cellulase produced in this manner could tentatively be used to hydrolyze fresh fiber sludge to obtain medium suitable for production of BC in the same biorefinery.
Proposed roadmap to stepwise integration of genetics in family medicine and clinical research
Authors: Houwink Elisa JF , Sollie Annet W , Numans Mattijs E , Cornel Martina C
Published: February 2013

We propose A step‐by‐step roadmap to integrate genetics in the Electronic Patient Record in Family Medicine and clinical research. This could make urgent operationalization of readily available genetic knowledge feasible in clinical research and consequently improved medical care.

Improving genomic literacy by training and education is needed first. The second step is the improvement of the possibilities to register the family history in such a way that queries can identify patients at risk. Adding codes to the ICPC chapters “A21 Personal/family history of malignancy” and “A99 Disease carrier not described further” is proposed. Multidisciplinary guidelines for referral must be unambiguous. Electronical patient records need possibilities to add (new) family history information, including links between individuals who are family members. Automatic alerts should help general practitioners to recognize patients at risk who satisfy referral criteria. We present a familial breast cancer case with a BRCA1 mutation as an example.
Does land abandonment decrease species richness and abundance of plants and animals in Mediterranean pastures, arable lands and permanent croplands?
Authors: Plieninger Tobias , Gaertner Mirijam , Hui Cang , Huntsinger Lynn
Published: February 2013

Background

Obscured by the more prevalent discussion of intensification and expansion of agricultural land, the impacts of the abandonment of many grasslands and croplands of the world in recent decades have received limited attention in the literature. Land abandonment is a common phenomenon in the Mediterranean Basin, a global biodiversity hotspot, but little is known about the impacts of this process on biodiversity. To upscale existing case-study insight to a Pan-Mediterranean level, we plan to perform a systematic review of the effects of land abandonment on plant and animal richness and abundance in pastures, arable lands and permanent crops of the Mediterranean Basin. In particular, we ask (1) which taxonomic groups are most affected by land abandonment; (2) whether different spatial and temporal scales of studies influence species richness and abundance outcomes; (3) whether previous land use and current anthropogenic impacts on abandoned lands determine differences in the number and abundance of species; and (4) whether landscape context modifies the impacts.

Methods

Our review will be based on searching scientific databases as well as the internet for empirical studies on the effects of land abandonment on biodiversity in the Mediterranean Basin. Study selection follows a three-stage process and includes study quality assessment. Extracted data will be synthesized through meta-analysis. Results will be presented in the form of a quantitative (e.g. figures indicating effect sizes, tables summarizing heterogeneity statistics) and narrative synthesis. Additional aspects will be addressed through meta-regression and sub-group analyses.
Developmental Programming: Postnatal Steroids Complete Prenatal Steroid Actions to Differentially Organize the GnRH Surge Mechanism and Reproductive Behavior in Female Sheep
Authors: Jackson Leslie M. , Mytinger Andrea , Roberts Eila K. , Lee Theresa M. , Foster Douglas L. , Padmanabhan Vasantha , Jansen Heiko T.
Published: February 2013

In female sheep, estradiol (E2) stimulates the preovulatory GnRH/LH surge and receptive behavior, whereas progesterone blocks these effects. Prenatal exposure to testosterone disrupts both the positive feedback action of E2 and sexual behavior although the mechanisms remain unknown. The current study tested the hypothesis that both prenatal and postnatal steroids are required to organize the surge and sex differences in reproductive behavior. Our approach was to characterize the LH surge and mating behavior in prenatally untreated (Control) and testosterone-treated (T) female sheep subsequently exposed to one of three postnatal steroid manipulations: endogenous E2, excess E2 from a chronic implant, or no E2 due to neonatal ovariectomy (OVX). All females were then perfused at the time of the expected surge and brains processed for estrogen receptor and Fos immunoreactivity. None of the T females exposed postnatally to E2 exhibited an E2-induced LH surge, but a surge was produced in five of six T/OVX and all Control females. No surges were produced when progesterone was administered concomitantly with E2. All Control females were mounted by males, but significantly fewer T females were mounted by a male, including the T/OVX females that exhibited LH surges. The percentage of estrogen receptor neurons containing Fos was significantly influenced in a brain region-, developmental stage-, and steroid-specific fashion by testosterone and E2 treatments. These findings support the hypothesis that the feedback controls of the GnRH surge are sensitive to programming by prenatal and postnatal steroids in a precocial species.
GH-Releasing Hormone Induces Cardioprotection in Isolated Male Rat Heart via Activation of RISK and SAFE Pathways
Authors: Penna Claudia , Settanni Fabio , Tullio Francesca , Trovato Letizia , Pagliaro Pasquale , Alloatti Giuseppe , Ghigo Ezio , Granata Riccarda
Published: February 2013

GHRH stimulates GH synthesis and release from the pituitary and exerts direct effects in extrapituitary tissues. We have previously shown that pretreatment with GHRH reduces cardiomyocyte apoptosis and improves heart function in isolated rat hearts subjected to ischemia/reperfusion (I/R). Here, we determined whether GHRH given at reperfusion reduces myocardial reperfusion injury and investigated the molecular mechanisms involved in GHRH effects. Isolated rat hearts subjected to I/R were treated at the onset of reperfusion with: 1) GHRH; 2) GHRH+GHRH antagonist JV-1-36; 3) GHRH+mitochondrial ATP-dependent potassium channel inhibitor 5-hydroxydecanoate; 4) GHRH+mitochondrial permeability transition pore opener atractyloside; 5) GHRH+ phosphoinositide 3-kinase/Akt inhibitor Wortmannin (WM); and 6) GHRH+signal transducer and activator of transcription-3 inhibitor tyrphostin-AG490 (AG490). GHRH reduced infarct size at the end of reperfusion and reverted contractility dysfunction in I/R hearts. These effects were inhibited by either JV-1-36, 5-hydroxydecanoate, atractylosid, WM, or AG490. Western blot analysis on left ventricles showed GHRH-induced phosphorylation of either the reperfusion injury salvage kinases (RISK), phosphoinositide 3-kinase/Akt, ERK1/2, and glycogen synthase kinase-3β or signal transducer and activator of transcription-3, as part of the survivor activating factor enhancement (SAFE) pathway. GHRH-induced activation of RISK and SAFE pathways was blocked by JV-1-36, WM, and AG490. Furthermore, GHRH increased the phosphorylation of endothelial nitric oxide synthase and AMP-activated protein kinase and preserved postischemic nicotinamide adenine dinucleotide (NAD+) levels. These results suggest that GHRH protects the heart from I/R injury through receptor-mediated mechanisms, leading to activation of RISK and SAFE pathways, which converge on mitochondria and possibly on AMP-activated protein kinase.
Excess Androgen During Puberty Disrupts Circadian Organization in Female Rats
Authors: Sellix Michael T. , Murphy Zachary C. , Menaker Michael
Published: February 2013

Circadian clocks have been described in each tissue of the hypothalamo-pituitary-ovarian axis. Although a role for the clock in the timing of ovulation is indicated, the impact of diseases that disrupt fertility on clock function or the clocks' role in the etiology of these pathologies has yet to be fully appreciated. Polycystic ovary syndrome (PCOS) is a particularly devastating endocrinopathy, affecting approximately 10% of women at childbearing age. Common features of PCOS are a polycystic ovary, amenorrhea, and excess serum androgen. Approximately 40% of these women have metabolic syndrome, including hyperinsulinemia, dyslipidemia, and hyperleptinemia. It has been suggested that excess androgen is a critical factor in the etiology of PCOS. We have examined the effects of androgen excess during puberty on the phase of circadian clocks in tissues of the metabolic and hypothalamo-pituitary-ovarian axes. Female period1-luciferase (per1-luc) rats were exposed to androgen (5α-dihydrotestosterone [DHT]) or placebo for 4-6 weeks (short term) or 9-15 weeks (long term). As expected, DHT-treated animals gained more weight than controls and had disrupted estrous cycles. At the end of treatment, tissues, including the liver, lung, kidney, white adipose, cornea, pituitary, oviduct, and ovarian follicles, were cultured, and per1-luc expression in each was recorded. Analysis of per1-luc expression revealed that DHT exposure increased phase distribution of multiple oscillators, including ovarian follicles, liver, and adipose, and altered phase synchrony between animals. These data suggest that excess androgen during puberty, a common feature of PCOS, negatively affects internal circadian organization in both the reproductive and metabolic axes.
Hepatic Growth Hormone Resistance After Acute Injury
Authors: Corrick Ryan M. , Li Li , Frank Stuart J. , Messina Joseph L.
Published: February 2013

Severe injury and infection are often followed by accelerated protein catabolism and acute insulin resistance. This results in several effects that complicate and prolong recovery, including weakness, immobility, impaired wound healing, and organ dysfunction. Recent studies have demonstrated the development of GH resistance during severe inflammation, providing a potential mechanism for the protein loss that follows injury and infection. To understand this GH resistance, we recently developed a murine model of acute injury. Mice were subjected to soft-tissue injury, alone or combined with hemorrhage, and injected iv with GH 30, 60, or 90 minutes later. Hepatic GH signaling was measured via Western analysis. GH-induced signal transducer and activator of transcription 5 phosphorylation was decreased immediately after completion of the trauma procedure, and at 30 and 60 minutes, but further decreased by 90 minutes after trauma. Combined trauma and hemorrhage resulted in severely decreased GH-induced signal transducer and activator of transcription 5 phosphorylation compared with trauma alone, and this was true at all time points studied. Western analysis revealed an apparent decrease in the molecular weight of the hepatic GH receptor (GHR) after trauma and hemorrhage, but not trauma alone. Additional studies determined that the hemorrhage-induced decrease in receptor size was not due to changes in GHR N-linked glycosylation. These results suggest that GH sensitivity is rapidly impaired after acute injury and that trauma combined with hemorrhage results in a more severe form of GH resistance resulting from alteration or inactivation of hepatic GHR.
Ghrelin Restoration of Function In Vitro in Somatotropes from Male Mice Lacking the Janus Kinase (JAK)-Binding Site of the Leptin Receptor
Authors: Syed Mohsin , Cozart Michael , Haney Anessa C. , Akhter Noor , Odle Angela K. , Allensworth-James Melody , Crane Christopher , Syed Farhan M. , Childs Gwen V.
Published: February 2013

Deletion of the signaling domain of leptin receptors selectively in somatotropes, with Cre-loxP technology, reduced the percentage of immunolabeled GH cells and serum GH. We hypothesized that the deficit occurred when leptin's postnatal surge failed to stimulate an expansion in the cell population. To learn more about the deficiency in GH cells, we tested their expression of GHRH receptors and GH mRNA and the restorative potential of secretagogue stimulation in vitro. In freshly plated dissociated pituitary cells from control male mice, GHRH alone (0.3 nM) increased the percentage of immunolabeled GH cells from 27 ± 0.05% (vehicle) to 42 ± 1.8% (P < .002) and the secretion of GH 1.8–3×. Deletion mutant pituitary cells showed a 40% reduction in percentages of immunolabeled GH cells (16.7 ± 0.4%), which correlated with a 47% reduction in basal GH levels (50 ng/mL control; 26.7 ng/mL mutants P = .01). A 50% reduction in the percentage of mutant cells expressing GHRH receptors (to 12%) correlated with no or reduced responses to GHRH. Ghrelin alone (10 nM) stimulated more GH cells in mutants (from 16.7–23%). When added with 1–3 nM GHRH, ghrelin restored GH cell percentages and GH secretion to levels similar to those of stimulated controls. Counts of somatotropes labeled for GH mRNA confirmed normal percentages of somatotropes in the population. These discoveries suggest that leptin may optimize somatotrope function by facilitating expression of membrane GHRH receptors and the production or maintenance of GH stores.
Phosphorylation of Threonine 333 Regulates Trafficking of the Human sst5 Somatostatin Receptor
Authors: Petrich Aline , Mann Anika , Kliewer Andrea , Nagel Falko , Strigli Anne , Märtens Jan Carlo , Pöll Florian , Schulz Stefan
Published: February 2013

The frequent overexpression of the somatostatin receptors sst2 and sst5 in neuroendocrine tumors provides the molecular basis for therapeutic application of novel multireceptor somatostatin analogs. Although the phosphorylation of the carboxyl-terminal region of the sst2 receptor has been studied in detail, little is known about the agonist-induced regulation of the human sst5 receptor. Here, we have generated phosphosite-specific antibodies for the carboxyl-terminal threonines 333 (T333) and 347 (T347), which enabled us to selectively detect either the T333-phosphorylated or the T347-phosphorylated form of sst5. We show that agonist-mediated phosphorylation occurs at T333, whereas T347 is constitutively phosphorylated in the absence of agonist. We further demonstrate that the multireceptor somatostatin analog pasireotide and the sst5-selective ligand L-817,818 but not octreotide or KE108 were able to promote a detectable T333 phosphorylation. Interestingly, BIM-23268 was the only sst5 agonist that was able to stimulate T333 phosphorylation to the same extent as natural somatostatin. Agonist-induced T333 phosphorylation was dose-dependent and selectively mediated by G protein-coupled receptor kinase 2. Similar to that observed for the sst2 receptor, phosphorylation of sst5 occurred within seconds. However, unlike that seen for the sst2 receptor, dephosphorylation and recycling of sst5 were rapidly completed within minutes. We also identify protein phosphatase 1γ as G protein-coupled receptor phosphatase for the sst5 receptor. Together, we provide direct evidence for agonist-selective phosphorylation of carboxyl-terminal T333. In addition, we identify G protein-coupled receptor kinase 2-mediated phosphorylation and protein phosphatase 1γ-mediated dephosphorylation of T333 as key regulators of rapid internalization and recycling of the human sst5 receptor.
The Politics and Strategy of Industry Self-Regulation: The Pharmaceutical Industry's Principles for Ethical Direct-to-Consumer Advertising as a Deceptive Blocking Strategy
Authors: Arnold Denis G. , Oakley James L.
Published: February 2013

As the pharmaceutical industry lobbies European regulators to permit direct-to-consumer advertising (DTCA) of prescription drugs in the European Union, we found that five leading companies violated industry-developed and -promulgated standards for ethical advertising in the United States. Utilizing multiple data sources and methods, we demonstrate a consistent failure by companies that market erectile dysfunction drugs to comply with the industry's guiding principles for ethical DTCA over a four-year period despite pledges of compliance by company leaders. Noncompliance resulted in children being exposed to sexually themed promotional messages more than 100 billion times. We argue that the guidelines are a coordinated effort by the industry to prevent unwanted federal regulation, and we introduce the concept of a blocking strategy to explain company behavior and to advance theoretical understanding of firms' public affairs strategies. We recommend policy responses to prevent deceptive practices, protect children from adult content, and promote genuine health care education.
Generic Drug Names and Social Welfare
Authors: Lobo Félix , Feldman Roger
Published: February 2013

This article studies how well International Nonproprietary Names (INNs), the “generic” names for pharmaceuticals, address the problems of imperfect information. Left in private hands, the identification of medicines leads to confusion and errors. Developed in the 1950s by the World Health Organization, INNs are a common, global, scientific nomenclature designed to overcome this failure. Taking stock after sixty years, we argue that the contribution of INNs to social welfare is paramount. They enhance public health by reducing errors and improving patient safety. They also contribute to economic efficiency by creating transparency as the foundation of competitive generic drug markets, reducing transaction costs, and favoring trade. The law in most countries requires manufacturers to designate pharmaceuticals with INNs in labeling and advertising. Generic substitution is also permitted or mandatory in many countries. But not all the benefits of INNs are fully realized because prescribers may not use them. We advocate strong incentives or even legally binding provisions to extend the use of INNs by prescribing physicians and dispensing pharmacists, but we do not recommend replacing brand names entirely with INNs. Instead, we propose dual use of brand names and INNs in prescribing, as in drug labeling.
Provincial Drug Plan Officials' Views of the Canadian Drug Safety System
Authors: Lexchin Joel , Wiktorowicz Mary , Moscou Kathy , Eggertson Laura
Published: February 2013

The Canadian constitution divides the responsibility for pharmaceuticals between the federal and provincial governments. While the provincial governments are responsible for establishing public formularies, the majority of the safety and efficacy information that the provinces use comes from the federal government. We interviewed drug plan officials from eight of the ten provinces and two of three territories regarding their views on the Canadian drug safety system. Here we report on the following categories: the federal drug approval system; the strengths and weaknesses of the federal system of postmarket pharmaceutical safety (i.e., pharmacosurveillance); resources available to support provincial formulary decision making; provincial roles in pharmacosurveillance; how the drug safety system could be improved; and the role of the Drug Safety and Effectiveness Network, a recently established virtual network designed to connect researchers throughout Canada who conduct postmarket drug research. Next, we place the Canadian system within an international context by comparing informational asymmetry between government institutions in the United States and the European Union and by looking at how institutions support each other's roles in sharing information and in jointly developing policy through the International Conference on Harmonization. Finally, we draw on international experiences and suggest potential solutions to the concerns that our key informants have identified.
How to Think Clearly About Medicare Administrative Costs: Data Sources and Measurement
Authors: Sullivan Kip
Published: February 2013

The Centers for Medicare and Medicaid Services (CMS) annually publishes two measures of Medicare's administrative expenditures. One of these appears in the reports of the Medicare Boards of Trustees and the other in the National Health Expenditure Accounts (NHEA). The latest trustees' report indicates Medicare's administrative expenditures are 1 percent of total Medicare spending, while the latest NHEA indicates the figure is 6 percent. The debate about Medicare's administrative expenditures, which emerged several years ago, reflects widespread confusion about these data. Critics of Medicare argue that the official reports on Medicare's overhead ignore or hide numerous types of administrative spending, such as the cost of collecting taxes and Part B premiums. Defenders of Medicare claim the official statistics are accurate. But participants on both sides of this debate fail to cite the official documents and do not analyze CMS's methodology. This article examines the controversy over the methodology CMS uses to calculate the trustees' and NHEA's measures and the sources of confusion and ignorance about them. It concludes with a discussion of how the two measures should be used.
Basal aurora kinase B activity is sufficient for histone H3 phosphorylation in prophase
Authors: Le Ly-Thuy-Tram , Vu Hong-Lien , Nguyen Chi-Hung , Molla Annie
Published: February 2013

Histone H3 phosphorylation is the hallmark of mitosis deposited by aurora kinase B. Benzo[e]pyridoindoles are a family of potent, broad, ATP-competitive aurora kinase inhibitors. However, benzo[e]pyridoindole C4 only inhibits histone H3 phosphorylation in prophase but not in metaphase. Under the C4 treatment, the cells enter into mitosis with dephosphorylated histone H3, assemble chromosomes normally and progress to metaphase, and then to anaphase. C4 also induces lagging chromosome in anaphase but we demonstrated that these chromosome compaction defects are not related to the absence of H3 phosphorylation in prophase. As a result of C4 action, mitosis lasts longer and the cell cycle is slowed down.

We reproduced the mitotic defects with reduced concentrations of potent pan aurora kinase as well as with a specific aurora B ATP-competitive inhibitor; we therefore propose that histone H3 phosphorylation and anaphase chromosome compaction involve the basal activity of aurora kinase B. Our data suggest that aurora kinase B is progressively activated at mitosis entry and at anaphase onset. The full activation of aurora kinase B by its partners, in prometaphase, induces a shift in the catalytic domain of aurora B that modifies its affinity for ATP. These waves of activation/deactivation of aurora B correspond to different conformations of the chromosomal complex revealed by FRAP.

The presence of lagging chromosomes may have deleterious consequences on the daughter cells and, unfortunately, the situation may be encountered in patients receiving treatment with aurora kinase inhibitors.
Analysis of the shape fluctuations of reconstituted membranes using GUVs made from lipid extracts of invertebrates
Authors: Bouvrais Hélène , Holmstrup Martin , Westh Peter , Ipsen John H.
Published: February 2013

Changes in the physical properties of the lipid matrix of cell membranes have repeatedly been proposed to underlie stresses associated with e.g. drought, cold and xenobiotics. Therefore, the ability to experimentally monitor such properties is central to the fundamental physiological understanding of adaptive changes. Here, we test the analysis of shape fluctuations in membranes composed of lipid extracts from two soil invertebrates, and show that theories and experimental approaches previously developed for simpler liposomes may be applied directly to reconstituted membrane lipids. Specifically, we show how the bending rigidity of giant unilamellar liposomes of lipid extracts can be determined precisely. We suggest that future measurements of this parameter could elucidate mechanisms of adaptive processes such as changes in lipid composition and accumulation of protective osmolytes.