bluuebunny/crossref_metadata_2025 huggingface dataset:
Systemic treatment of liver metastases from colorectal cancer
Authors: Stein Alexander , Schmoll Hans-Joachim
Published: March 2013

Patients presenting with synchronous or metachronous colorectal cancer liver metastases (CLM) should be evaluated for multimodal management with curative intent. Preoperative systemic chemotherapy shows beneficial impact on adjuvant progression-free survival and also borderline on overall survival, without significantly increasing initially R0 resectable patients postoperative complication rates. Postoperative chemotherapy recommended based on the perioperative trial experience for those patients achieving at least stable disease during preoperative chemotherapy, or based on the adjuvant trials for patients receiving upfront resection. ‘Borderline’ resectable CLM, preoperative chemotherapy plays an important role in both in achievement of a resectable status and improvement of prognosis. Recent 4 drug combinations demonstrated response rates up to 80% even for advanced disease and are thus promising regimens for further evaluation in patients with resectable or unresectable liver-limited (+/- lung) disease.
Manufacturing a Messiah
Authors: Mocarski Richard , Billings Andrew C.
Published: March 2013

This monograph analyzes how Nike and the athlete himself jointly commodified the public persona of LeBron James, with James cast in two normative narratives: (a) the Messiah and (b) hegemonic masculinity, stripping James of his Blackness and making him identifiable to a mainstream audience. The open-ended configuration of these narratives allows for “pivot points” in James’ life. Real-life developments fold into the construction of James, mitigating damage and shaping narratives in the process. New avenues of research in critical celebrity-branding analysis focus on areas such as the cross sections of endorsements and social media as well as the process of how mediated narratives normalize subjectivities.
In situ hybridization protocol for enhanced detection of gene expression in the planarian Schmidtea mediterranea
Authors: King Ryan S , Newmark Phillip A
Published: March 2013

Background

The freshwater planarian Schmidtea mediterranea has emerged as a powerful model for studies of regenerative, stem cell, and germ cell biology. Whole-mount in situ hybridization (WISH) and whole-mount fluorescent in situ hybridization (FISH) are critical methods for determining gene expression patterns in planarians. While expression patterns for a number of genes have been elucidated using established protocols, determining the expression patterns for particularly low-abundance transcripts remains a challenge.

Results

We show here that a short bleaching step in formamide dramatically enhances signal intensity of WISH and FISH. To further improve signal sensitivity we optimized blocking conditions for multiple anti-hapten antibodies, developed a copper sulfate quenching step that virtually eliminates autofluorescence, and enhanced signal intensity through iterative rounds of tyramide signal amplification. For FISH on regenerating planarians, we employed a heat-induced antigen retrieval step that provides a better balance between permeabilization of mature tissues and preservation of regenerating tissues. We also show that azide most effectively quenches peroxidase activity between rounds of development for multicolor FISH experiments. Finally, we apply these modifications to elucidate the expression patterns of a few low-abundance transcripts.

Conclusion

The modifications we present here provide significant improvements in signal intensity and signal sensitivity for WISH and FISH in planarians. Additionally, these modifications might be of widespread utility for whole-mount FISH in other model organisms.
Gene rearrangements in hormone receptor negative breast cancers revealed by mate pair sequencing
Authors: Jiao Xiang , Hooper Sean D , Djureinovic Tatjana , Larsson Chatarina , Wärnberg Fredrik , Tellgren-Roth Christian , Botling Johan , Sjöblom Tobias
Published: March 2013

Background

Chromosomal rearrangements in the form of deletions, insertions, inversions and translocations are frequently observed in breast cancer genomes, and a subset of these rearrangements may play a crucial role in tumorigenesis. To identify novel somatic chromosomal rearrangements, we determined the genome structures of 15 hormone-receptor negative breast tumors by long-insert mate pair massively parallel sequencing.

Results

We identified and validated 40 somatic structural alterations, including the recurring fusion between genes DDX10 and SKA3 and translocations involving the EPHA5 gene. Other rearrangements were found to affect genes in pathways involved in epigenetic regulation, mitosis and signal transduction, underscoring their potential role in breast tumorigenesis. RNA interference-mediated suppression of five candidate genes (DDX10, SKA3, EPHA5, CLTC and TNIK) led to inhibition of breast cancer cell growth. Moreover, downregulation of DDX10 in breast cancer cells lead to an increased frequency of apoptotic nuclear morphology.

Conclusions

Using whole genome mate pair sequencing and RNA interference assays, we have discovered a number of novel gene rearrangements in breast cancer genomes and identified DDX10, SKA3, EPHA5, CLTC and TNIK as potential cancer genes with impact on the growth and proliferation of breast cancer cells.
Association of decreased mitochondrial DNA content with the progression of colorectal cancer
Authors: Cui HaiHong , Huang Ping , Wang ZhiJing , Zhang YunXin , Zhang ZhenHua , Xu Wei , Wang XiaoPeng , Han Ying , Guo XiaoMing
Published: March 2013

Background

Experimental data suggest that mitochondria is involved in tumorigenesis. However, little is known about the qualitative and quantitative changes of mtDNA in colorectal cancer tissues. We therefore conducted possible correlations of the mitochondrial DNA (mtDNA) copy number in colorectal cancer (CRC) with clinical and pathological findings and CRC prognosis.

Methods

mtDNA copy numbers in CRC cancer tissue and adjacent non-cancerous tissue samples were measured using quantitative real-time polymerase chain reaction analyses from 60 patients admitted to our hospital. We examined the correlation of mtDNA copy numbers and clinicopathologic parameters of CRC patients. The correlation between mtDNA copy number and three-year survival was analyzed.

Results

The mtDNA copy number was lower in CRC tissue compared with the corresponding non-cancerous colorectal tissue (mean: 108.60 ± 20.11 vs. 153.68 ± 25.72) and was significantly correlated with lymph-node metastasis. Patients with a lower mtDNA copy number tended to have lower 3-year survival than patients with a higher mtDNA copy number assessed by Kaplan–Meier curves, but the correlation was not significant (overall survival, 63.0 vs 83%).

Conclusions

These results suggest that a reduced copy number of mtDNA is correlated with malignant potential in CRC.
Uptake of a web-based oncology protocol system: how do cancer clinicians use eviQ cancer treatments online?
Authors: Langton Julia M , Pesa Nicole , Rushton Shelley , Ward Robyn L , Pearson Sallie-Anne
Published: March 2013

Background

The use of computerized systems to support evidence-based practice is commonplace in contemporary medicine. Despite the prolific use of electronic support systems there has been relatively little research on the uptake of web-based systems in the oncology setting. Our objective was to examine the uptake of a web-based oncology protocol system (http://www.eviq.org.au) by Australian cancer clinicians.

Methods

We used web-logfiles and Google Analytics to examine the characteristics of eviQ registrants from October 2009-December 2011 and patterns of use by cancer clinicians during a typical month.

Results

As of December 2011, there were 16,037 registrants; 85% of whom were Australian health care professionals. During a typical month 87% of webhits occurred in standard clinical hours (08:00 to 18:00 weekdays). Raw webhits were proportional to the size of clinician groups: nurses (47% of Australian registrants), followed by doctors (20%), and pharmacists (14%). However, pharmacists had up to three times the webhit rate of other clinical groups. Clinicians spent five times longer viewing chemotherapy protocol pages than other content and the protocols viewed reflect the most common cancers: lung, breast and colorectal.

Conclusions

Our results demonstrate eviQ is used by a range of health professionals involved in cancer treatment at the point-of-care. Continued monitoring of electronic decision support systems is vital to understanding how they are used in clinical practice and their impact on processes of care and patient outcomes.
Optimizing heterologous protein production in the periplasm of E. coli by regulating gene expression levels
Authors: Schlegel Susan , Rujas Edurne , Ytterberg Anders Jimmy , Zubarev Roman A , Luirink Joen , de Gier Jan-Willem
Published: March 2013

Background

In Escherichia coli many heterologous proteins are produced in the periplasm. To direct these proteins to the periplasm, they are equipped with an N-terminal signal sequence so that they can traverse the cytoplasmic membrane via the protein-conducting Sec-translocon. For poorly understood reasons, the production of heterologous secretory proteins is often toxic to the cell thereby limiting yields. To gain insight into the mechanism(s) that underlie this toxicity we produced two secretory heterologous proteins, super folder green fluorescent protein and a single-chain variable antibody fragment, in the Lemo21(DE3) strain. In this strain, the expression intensity of the gene encoding the target protein can be precisely controlled.

Results

Both SFGFP and the single-chain variable antibody fragment were equipped with a DsbA-derived signal sequence. Producing these proteins following different gene expression levels in Lemo21(DE3) allowed us to identify the optimal expression level for each target gene. Too high gene expression levels resulted in saturation of the Sec-translocon capacity as shown by hampered translocation of endogenous secretory proteins and a protein misfolding/aggregation problem in the cytoplasm. At the optimal gene expression levels, the negative effects of the production of the heterologous secretory proteins were minimized and yields in the periplasm were optimized.

Conclusions

Saturating the Sec-translocon capacity can be a major bottleneck hampering heterologous protein production in the periplasm. This bottleneck can be alleviated by harmonizing expression levels of the genes encoding the heterologous secretory proteins with the Sec-translocon capacity. Mechanistic insight into the production of proteins in the periplasm is key to optimizing yields in this compartment.
The 3.5 ångström X−ray structure of the human connexin26 gap junction channel is unlikely that of a fully open channel
Authors: Zonta Francesco , Polles Guido , Sanasi Maria Federica , Bortolozzi Mario , Mammano Fabio
Published: March 2013

The permeability of gap junction channels to metabolites, and not simply to small inorganic ions, is likely to play an important role in development, physiology as well as in etiology of several diseases. Here, we combined dual patch clamp and fluorescence imaging techniques with molecular dynamics (MD) simulations to investigate the permeation of calcein, a relatively large fluorescent tracer (MW 622 Da) through homomeric gap junction channels formed by wild type human connexin26 (hCx26wt) protomers. Our experimental data indicate that the unitary flux of calcein driven by a 125 μM concentration difference is J
pore = 226 molecule/s per channel. In the light of Eyring transition state theory adapted for the liquid phase, this value corresponds to an energy barrier of ~20 k

B

T (where k

B
is the Boltzmann constant and T is absolute temperature). The barrier predicted by our MD simulations, based on the 3.5 Å X–ray structural model of the hCx26wt gap junction channel, is ~45 k

B

T. The main contributions to the energetics of calcein permeation originated from the interaction between the permeating molecule and the charged aminoacids lining the channel pore. Assigning a fake zero total charge to the calcein molecule yielded a value for the barrier height compatible with the experimental data. These results can be accounted for by two different (although not mutually exclusive) hypotheses: (1) the X–ray model of the hCx26wt gap junction channel is not representative of a fully open state; (2) post translational modifications affecting the hCx26wt protein in our expression system differed from the modifications undergone by the proteins in the conditions used to obtain the crystal structure. Hypothesis (1) is compatible with data indicating that, only 10% or less of the channels forming a gap junction plaque are in the open state, and therefore the averaging procedure intrinsic in the generation of the crystal structure data more closely reflects that of a closed channel. Hypothesis (2) is compatible with recent mass spectrometry data and implies that the charge of several amino acid side chains may have been altered, thus modifying substantially the permeation properties of the channels in living cells.
Immobilization of pamidronic acids on the nanotube surface of titanium discs and their interaction with bone cells
Authors: Koo Tae-Hyung , Borah Jyoti S , Xing Zhi-Cai , Moon Sung-Mo , Jeong Yongsoo , Kang Inn-Kyu
Published: March 2013

Self-assembled layers of vertically aligned titanium nanotubes were fabricated on a Ti disc by anodization. Pamidronic acids (PDAs) were then immobilized on the nanotube surface to improve osseointegration. Wide-angle X-ray diffraction, X-ray photoelectron microscopy, and scanning electron microscopy were employed to characterize the structure and morphology of the PDA-immobilized TiO2nanotubes. Thein vitrobehavior of osteoblast and osteoclast cells cultured on an unmodified and surface-modified Ti disc was examined in terms of cell adhesion, proliferation, and differentiation. Osteoblast adhesion, proliferation, and differentiation were improved substantially by the topography of the TiO2nanotubes, producing an interlocked cell structure. PDA immobilized on the TiO2nanotube surface suppressed the viability of the osteoclasts and reduced their bone resorption activity.
Low serum eicosapentaenoic acid / arachidonic acid ratio in male subjects with visceral obesity
Authors: Inoue Kana , Kishida Ken , Hirata Ayumu , Funahashi Tohru , Shimomura Iichiro
Published: March 2013

Background

Visceral fat accumulation is caused by over-nutrition and physical inactivity. Excess accumulation of visceral fat associates with atherosclerosis. Polyunsaturated fatty acids have an important role in human nutrition, but imbalance of dietary long-chain polyunsaturated fatty acids, especially low eicosapentaenoic acid (EPA) / arachidonic acid (AA) ratio, is associated with increased risk of cardiovascular disease. The present study investigated the correlation between EPA, docosahexaenoic acid (DHA), AA parameters and clinical features in male subjects.

Findings

The study subjects were 134 Japanese with diabetes, hypertension and/or dyslipidemia who underwent measurement of visceral fat area (eVFA) by the bioelectrical impedance method and serum levels of EPA, DHA and AA. EPA/AA ratio correlated positively with age, and negatively with waist circumference and eVFA. Stepwise regression analysis demonstrated that age and eVFA correlated significantly and independently with serum EPA/AA ratio. Serum EPA/AA ratio, but not serum DHA/AA and (EPA+DHA)/AA ratios, was significantly lower in subjects with eVFA ≥100 cm2, compared to those with eVFA <100 cm2 (p=0.049). Subjects with eVFA ≥100 cm2 were significantly more likely to have the metabolic syndrome and history of cardiovascular diseases, compared to those with eVFA <100 cm2 (p<0.001, p=0.028, respectively).

Conclusions

Imbalance of dietary long-chain polyunsaturated fatty acids (low serum EPA/AA ratio) correlated with visceral fat accumulation in male subjects.

Clinical trial registration number

UMIN000002271
Omega-3 fatty acids, polymorphisms and lipid related cardiovascular disease risk factors in the Inuit population
Authors: Rudkowska Iwona , Ouellette Catherine , Dewailly Eric , Hegele Robert A , Boiteau Véronique , Dubé-Linteau Ariane , Abdous Belkacem , Proust Françoise , Giguère Yves , Julien Pierre , Château-Degat Marie-Ludivine , Vohl Marie-Claude
Published: March 2013

Background

Tissue concentrations of fatty acids (FAs) and genetic variations are well-known factors which affect the cardiovascular disease (CVD) risk. The objective was to examine whether the genetic variability of 20 candidate genes and red blood cells (RBCs) percentage of total n-3 polyunsaturated fatty acids (PUFA), a biomarker of dietary n-3 PUFA intake, modulate lipid related CVD risk factors in the Inuit population.

Methods

Data from the Qanuippitaa Nunavik Health Survey (n = 553) were analysed via multivariate regression models with 40 known polymorphisms, RBCs percentage of n-3 PUFA, and the interaction term to take into account the effect on plasma lipid and apolipoporotein levels.

Results

Individuals being heterozygotes for CETP C-4502T (rs183130) or G-971A (rs4783961) together with higher n-3 PUFA had lower triacylglycerol (TG) concentrations compared to homozygotes for the minor allele. Further, effects of a stronger beneficial association between n-3 PUFA in RBCs and plasma lipid parameters- including lower total cholesterol (TC), lower low-density lipoprotein cholesterol (LDL-C) or higher high-density lipoprotein cholesterol (HDL-C) concentrations- were associated with AGT M235T (rs699) TT genotype, CETP G-971A (rs4783961) AG genotype, T allele carriers of CETP C-4502T (rs183130), and T allele carriers of CETP Ile405Val (rs5882). In contrast, higher n-3 PUFA in RBCs were associated with adverse lipid profiles- including increased LDL-C, increased apolipoprotein B100 or decreased HDL-C concentrations- in G allele carriers of the APOA5 -3 A/G (rs651821), C allele carriers of APOA5 T-1131C (rs662799), G carriers of APOC3 SstI (rs5128) and G carriers of APOA4 Asn147Ser (rs5104).

Conclusion

Overall, these results suggest that percentage of total n-3 PUFA of RBCs are associated with lipids related CVD risk factors conferred by genetic variations in the Inuit population.
Lithium metasilicate and lithium disilicate nanomaterials: optical properties and density functional theory calculations
Authors: Alemi Abdolali , Khademinia Shahin , Joo Sang Woo , Dolatyari Mahboubeh , Bakhtiari Akbar
Published: March 2013

UV–vis and photoluminescence spectra of the hydrothermally synthesized crystalline lithium metasilicate (Li2SiO3) and lithium disilicate (Li2Si2O5) nanomaterials are studied. The intensity of the bands in the emission spectra increases with increasing reaction time in both compounds. The electronic band structure along with density of states calculated by the density functional theory (DFT) method indicates that Li2SiO3 and Li2Si2O5 have an indirect energy band gap of 4.575 and 4.776 eV respectively. The optical properties, including the dielectric, absorption, reflectivity, and energy loss spectra of the compounds, are calculated by DFT method and analyzed based on the electronic structures.
Vitamin K-dependent proteins GAS6 and Protein S and TAM receptors in patients of systemic lupus erythematosus: correlation with common genetic variants and disease activity
Authors: Recarte-Pelz Pedro , Tàssies Dolors , Espinosa Gerard , Hurtado Begoña , Sala Núria , Cervera Ricard , Reverter Joan Carles , de Frutos Pablo García
Published: March 2013

Introduction

Growth arrest-specific gene 6 protein (GAS6) and protein S (ProS) are vitamin K-dependent proteins present in plasma with important regulatory functions in systems of response and repair to damage. They interact with receptor tyrosine kinases of the Tyro3, Axl and MerTK receptor tyrosine kinase (TAM) family, involved in apoptotic cell clearance (efferocytosis) and regulation of the innate immunity. TAM-deficient mice show spontaneous lupus-like symptoms. Here we tested the genetic profile and plasma levels of components of the system in patients with systemic lupus erythematosus (SLE), and compare them with a control healthy population.

Methods

Fifty SLE patients and 50 healthy controls with matched age, gender and from the same geographic area were compared. Genetic analysis was performed in GAS6 and the TAM receptor genes on SNPs previously identified. The concentrations of GAS6, total and free ProS, and the soluble forms of the three TAM receptors (sAxl, sMerTK and sTyro3) were measured in plasma from these samples.

Results

Plasma concentrations of GAS6 were higher and, total and free ProS were lower in the SLE patients compared to controls, even when patients on oral anticoagulant treatment were discarded. Those parameters correlated with SLE disease activity index (SLEDAI) score, GAS6 being higher in the most severe cases, while free and total ProS were lower. All 3 soluble receptors increased its concentration in plasma of lupus patients.

Conclusions

The present study highlights that the GAS6/ProS-TAM system correlates in several ways with disease activity in SLE. We show here that this correlation is affected by common polymorphisms in the genes of the system. These findings underscore the importance of mechanism of regulatory control of innate immunity in the pathology of SLE.
Endoplasmic reticulum stress induces PRNP prion protein gene expression in breast cancer
Authors: Déry Marc-André , Jodoin Julie , Ursini-Siegel Josie , Aleynikova Olga , Ferrario Cristiano , Hassan Saima , Basik Mark , LeBlanc Andréa C
Published: March 2013

Introduction

High prion protein (PrP) levels are associated with breast, colon and gastric cancer resistance to treatment and with a poor prognosis for the patients. However, little is known about the underlying molecular mechanism(s) regulating human PrP gene (PRNP) expression in cancers. Because endoplasmic reticulum (ER) stress is associated with solid tumors, we investigated a possible regulation of PRNP gene expression by ER stress.

Methods

Published microarray databases of breast cancer tissues and breast carcinoma cell lines were analyzed for PrP mRNA and ER stress marker immunoglobulin heavy chain binding protein (BiP) levels. Breast cancer tissue microarrays (TMA) were immunostained for BiP and PrP. Breast carcinoma MCF-7, MDA-MB-231, HS578T and HCC1500 cells were treated with three different ER stressors - Brefeldin A, Tunicamycin, Thapsigargin - and levels of PrP mRNA or protein assessed by RT-PCR and Western blot analyses. A human PRNP promoter-luciferase reporter was used to assess transcriptional activation by ER stressors. Site-directed mutagenesis identified the ER stress response elements (ERSE). Chromatin immunoprecipitation (ChIP) analyses were done to identify the ER stress-mediated transcriptional regulators. The role of cleaved activating transcription factor 6α (ΔATF6α) and spliced X-box protein-1 (sXBP1) in PRNP gene expression was assessed with over-expression or silencing techniques. The role of PrP protection against ER stress was assessed with PrP siRNA and by using Prnp null cell lines.

Results

We find that mRNA levels of BiP correlated with PrP transcript levels in breast cancer tissues and breast carcinoma cell lines. PrP mRNA levels were enriched in the basal subtype and were associated with poor prognosis in breast cancer patients. Higher PrP and BiP levels correlated with increasing tumor grade in TMA. ER stress was a positive regulator of PRNP gene transcription in MCF-7 cells and luciferase reporter assays identified one ER stress response element (ERSE) conserved among primates and rodents and three primate-specific ERSEs that regulated PRNP gene expression. Among the various transactivators of the ER stress-regulated unfolded protein response (UPR), ATF6α and XBP1 transactivated PRNP gene expression, but the ability of these varied in different cell types. Functionally, PrP delayed ER stress-induced cell death.

Conclusions

These results establish PRNP as a novel ER stress-regulated gene that could increase survival in breast cancers.
Phase III Randomized Study of Rituximab/Carmustine, Etoposide, Cytarabine, and Melphalan (BEAM) Compared With Iodine-131 Tositumomab/BEAM With Autologous Hematopoietic Cell Transplantation for Relapsed Diffuse Large B-Cell Lymphoma: Results From the BMT CTN 0401 Trial
Authors: Vose Julie M. , Carter Shelly , Burns Linda J. , Ayala Ernesto , Press Oliver W. , Moskowitz Craig H. , Stadtmauer Edward A. , Mineshi Shin , Ambinder Richard , Fenske Timothy , Horowitz Mary , Fisher Richard , Tomblyn Marcie
Published: March 2013

Purpose

This clinical trial evaluated standard-dose radioimmunotherapy with a chemotherapy-based transplantation regimen followed by autologous hematopoietic cell transplantation versus rituximab with the same regimen in patients with relapsed diffuse large B-cell lymphoma (DLBCL).

Patients and Methods

Patients with chemotherapy-sensitive persistent or relapsed DLBCL were randomly assigned to receive iodine-131 tositumomab (dosimetric dose of 5 mCi on day −19 and therapeutic dose of 0.75 Gy on day −12), carmustine 300 mg/m2(day −6), etoposide 100 mg/m2twice daily (days −5 to −2), cytarabine 100 mg/m2twice daily (days −5 to −2), and melphalan 140 mg/m2(day −1; B-BEAM) or rituximab 375 mg/m2on days −19 and −12 and the same chemotherapy regimen (R-BEAM).

Results

Two hundred twenty-four patients were enrolled, with 113 patients randomly assigned to R-BEAM and 111 patients assigned to B-BEAM. Two-year progression-free survival (PFS) rates, the primary end point, were 48.6% (95% CI, 38.6% to 57.8%) for R-BEAM and 47.9% (95% CI, 38.2% to 57%; P = .94) for B-BEAM, and the 2-year overall survival (OS) rates were 65.6% (95% CI, 55.3% to 74.1%) for R-BEAM and 61% (95% CI, 50.9% to 69.9%; P = .38) for B-BEAM. The 100-day treatment-related mortality rates were 4.1% (95% CI, 0.2% to 8.0%) for R-BEAM and 4.9% (95% CI, 0.8% to 9.0%; P = .97) for B-BEAM. The maximum mucositis score was higher in the B-BEAM arm (0.72) compared with the R-BEAM arm (0.31; P < .001).

Conclusion

The B-BEAM and R-BEAM regimens produced similar 2-year PFS and OS rates for patients with chemotherapy-sensitive relapsed DLBCL. No differences in toxicities other than mucositis were noted.
TCTEX1D4, a novel protein phosphatase 1 interactor: connecting the phosphatase to the microtubule network
Authors: Korrodi-Gregório Luís , Vieira Sandra I. , Esteves Sara L. C. , Silva Joana V. , Freitas Maria João , Brauns Ann-Kristin , Luers Georg , Abrantes Joana , Esteves Pedro J. , da Cruz e Silva Odete A. B. , Fardilha Margarida , da Cruz e Silva Edgar F.
Published: March 2013

Reversible phosphorylation plays an important role as a mechanism of intracellular control in eukaryotes. PPP1, a major eukaryotic Ser/Thr-protein phosphatase, acquires its specificity by interacting with different protein regulators, also known as PPP1 interacting proteins (PIPs). In the present work we characterized a physiologically relevant PIP in testis. Using a yeast two-hybrid screen with a human testis cDNA library, we identified a novel PIP of PPP1CC2 isoform, the T-complex testis expressed protein 1 domain containing 4 (TCTEX1D4) that has recently been described as a Tctex1 dynein light chain family member. The overlay assays confirm that TCTEX1D4 interacts with the different spliced isoforms of PPP1CC. Also, the binding domain occurs in the N-terminus, where a consensus PPP1 binding motif (PPP1BM) RVSF is present. The distribution of TCTEX1D4 in testis suggests its involvement in distinct functions, such as TGFβ signaling at the blood–testis barrier and acrosome cap formation. Immunofluorescence in human ejaculated sperm shows that TCTEX1D4 is present in the flagellum and in the acrosome region of the head. Moreover, TCTEX1D4 and PPP1 co-localize in the microtubule organizing center (MTOC) and microtubules in cell cultures. Importantly, the TCTEX1D4 PPP1BM seems to be relevant for complex formation, for PPP1 retention in the MTOC and movement along microtubules.

These novel results open new avenues to possible roles of this dynein, together with PPP1. In essence TCTEX1D4/PPP1C complex appears to be involved in microtubule dynamics, sperm motility, acrosome reaction and in the regulation of the blood–testis barrier.
Single-cell gene expression profiling reveals functional heterogeneity of undifferentiated human epidermal cells
Authors: Tan David W. M. , Jensen Kim B. , Trotter Matthew W. B. , Connelly John T. , Broad Simon , Watt Fiona M.
Published: March 2013

Human epidermal stem cells express high levels of β1 integrins, delta-like 1 (DLL1) and the EGFR antagonist LRIG1. However, there is cell-to-cell variation in the relative abundance of DLL1 and LRIG1 mRNA transcripts. Single-cell global gene expression profiling showed that undifferentiated cells fell into two clusters delineated by expression of DLL1 and its binding partner syntenin. The DLL1+ cluster had elevated expression of genes associated with endocytosis, integrin-mediated adhesion and receptor tyrosine kinase signalling. Differentially expressed genes were not independently regulated, as overexpression of DLL1 alone or together with LRIG1 led to the upregulation of other genes in the DLL1+ cluster. Overexpression of DLL1 and LRIG1 resulted in enhanced extracellular matrix adhesion and increased caveolin-dependent EGFR endocytosis. Further characterisation of CD46, one of the genes upregulated in the DLL1+ cluster, revealed it to be a novel cell surface marker of human epidermal stem cells. Cells with high endogenous levels of CD46 expressed high levels of β1 integrin and DLL1 and were highly adhesive and clonogenic. Knockdown of CD46 decreased proliferative potential and β1 integrin-mediated adhesion. Thus, the previously unknown heterogeneity revealed by our studies results in differences in the interaction of undifferentiated basal keratinocytes with their environment.
Intradermal adipocytes mediate fibroblast recruitment during skin wound healing
Authors: Schmidt Barbara A. , Horsley Valerie
Published: March 2013

Acute wound healing in the skin involves the communication of multiple cell types to coordinate keratinocyte and fibroblast proliferation and migration for epidermal and dermal repair. Many studies have focused on the interplay between hematopoietic cells, keratinocytes and fibroblasts during skin wound healing, yet the possible roles for other cell types within the skin, such as intradermal adipocytes, have not been investigated during this process. Here, we identify that adipocyte lineage cells are activated and function during acute skin wound healing. We find that adipocyte precursor cells proliferate and mature adipocytes repopulate skin wounds following inflammation and in parallel with fibroblast migration. Functional analysis of mice with defects in adipogenesis demonstrates that adipocytes are necessary for fibroblast recruitment and dermal reconstruction. These data implicate adipocytes as a key component of the intercellular communication that mediates fibroblast function during skin wound healing.
sfrp1 promotes cardiomyocyte differentiation in Xenopus via negative-feedback regulation of Wnt signalling
Authors: Gibb Natalie , Lavery Danielle L. , Hoppler Stefan
Published: March 2013

Wnt signalling is a key regulator of vertebrate heart development, yet it is unclear which specific Wnt signalling components are required to regulate which aspect of cardiogenesis. Previously, we identified Wnt6 as an endogenous Wnt ligand required for controlling heart muscle differentiation via canonical Wnt/β-catenin signalling. Here we show for the first time a requirement for an endogenous Wnt signalling inhibitor for normal heart muscle differentiation. Expression of sfrp1 is strongly induced in differentiating heart muscle. We show that sfrp1 is not only able to promote heart muscle differentiation but is also required for the formation of normal size heart muscle in the embryo. sfrp1 is functionally able to inhibit Wnt6 signalling and its requirement during heart development relates to relieving the cardiogenesis-restricting function of endogenous wnt6. In turn, we discover that sfrp1 expression in the heart is regulated by Wnt6 signalling, which for the first time indicates that sfrp genes can function as part of a Wnt negative-feedback regulatory loop. Our experiments indicate that sfrp1 controls the size of the differentiating heart muscle primarily by regulating cell fate within the cardiac mesoderm between muscular and non-muscular cell lineages. The cardiac mesoderm is therefore not passively patterned by signals from the surrounding tissue, but regulates its differentiation into muscular and non-muscular tissue using positional information from the surrounding tissue. This regulatory network might ensure that Wnt activation enables expansion and migration of cardiac progenitors, followed by Wnt inhibition permitting cardiomyocyte differentiation.
Floor plate-derived sonic hedgehog regulates glial and ependymal cell fates in the developing spinal cord
Authors: Yu Kwanha , McGlynn Sean , Matise Michael P.
Published: March 2013

Cell fate specification in the CNS is controlled by the secreted morphogen sonic hedgehog (Shh). At spinal cord levels, Shh produced by both the notochord and floor plate (FP) diffuses dorsally to organize patterned gene expression in dividing neural and glial progenitors. Despite the fact that two discrete sources of Shh are involved in this process, the individual contribution of the FP, the only intrinsic source of Shh throughout both neurogenesis and gliogenesis, has not been clearly defined. Here, we have used conditional mutagenesis approaches in mice to selectively inactivate Shh in the FP (ShhFP) while allowing expression to persist in the notochord, which underlies the neural tube during neurogenesis but not gliogenesis. We also inactivated Smo, the common Hh receptor, in neural tube progenitors. Our findings confirm and extend prior studies suggesting an important requirement for ShhFP in specifying oligodendrocyte cell fates via repression of Gli3 in progenitors. Our studies also uncover a connection between embryonic Shh signaling and astrocyte-mediated reactive gliosis in adults, raising the possibility that this pathway is involved in the development of the most common cell type in the CNS. Finally, we find that intrinsic spinal cord Shh signaling is required for the proper formation of the ependymal zone, the epithelial cell lining of the central canal that is also an adult stem cell niche. Together, our studies identify a crucial late embryonic role for ShhFP in regulating the specification and differentiation of glial and epithelial cells in the mouse spinal cord.
Sucos do bem: o desafio de crescer com a fruta na caixinha
Authors: Ferreira Daniela Abrantes , Ferreira Lucelena Abrantes , Mayer Verônica Feder
Published: March 2013

Marcos Leta fundou em 2007 a Sucos do bem, com a ideia de "fazer bebidas verdadeiras que proporcionam um momento divertido para pessoas jovens de espírito". O sucesso inicial traz o desafio da expansão da empresa para todo o país. Como fazer isso sem perder as características que contribuíram para o sucesso da iniciativa?DOI: 10.12660/gvcasosv2n1c1http://dx.doi.org/10.12660/gvcasosv2n1c1
Seguir carreira (ou não) em uma empresa familiar
Authors: Hehn Elisa Bornhausen Cunha , Lohn Deise , Pereira Daniela Cardoso , Mignoni Alissane , Marinho Sidnei Vieira
Published: March 2013

Carolina tem 27 anos e está indecisa se continua vinculada profissionalmente à empresa da família. As relações não são fáceis como parecem. Os âmbitos familiar e profissional se confundem, e o líder prioriza o papel de pai em detrimento do papel de gestor da empresa. Desde que Carolina assumiu a área financeira, constatou que seriam necessários reeducação de hábitos e maior comprometimento de todos. DOI: 10.12660/gvcasosv2n1c3http://dx.doi.org/10.12660/gvcasosv2n1c3
A internacionalização da marca Melissa (Notas de ensino)
Authors: Da Rocha Angela , De Almeida Moraes Sylvia Therezinha , Senra Melissa
Published: March 2013

DOI: 10.12660/gvcasosv2n1n6http://dx.doi.org/10.12660/gvcasosv2n1n6
Recuperação judicial de uma empresa aérea em crise econômico-financeira
Authors: Almeida Vinicio de Souza e , Romano Patrícia Ribeiro
Published: March 2013

A companhia Voe Mais é tradicional no mercado de transporte aéreo. Porém, vem sofrendo seguidos prejuízos. Como única alternativa para soerguimento da empresa, o diretor financeiro entrou com pedido de recuperação judicial, visando à proteção legal contra o arresto de aeronaves e a negociação de um plano de retomada com todos os credores. Será possível salvar a empresa?DOI: 10.12660/gvcasosv2n2c9http://dx.doi.org/10.12660/gvcasosv2n2c9
Concessões de termoelétricas no Brasil: investir ou não? (Notas de ensino)
Authors: Martits Luiz Augusto , Garcia Fábio Gallo , Amaral Júnior José Bento Carlos
Published: March 2013

DOI: 10.12660/gvcasosv2n2n7http://dx.doi.org/10.12660/gvcasosv2n2n7
Recuperação judicial de uma empresa aérea em crise econômico-financeira (Notas de ensino)
Authors: E Almeida Vinicio de Souza , Romano Patrícia Ribeiro
Published: March 2013

DOI: 10.12660/gvcasosv2n2n9http://dx.doi.org/10.12660/gvcasosv2n2n9
Variation in the Use of Diuretic Therapy for Infants With Bronchopulmonary Dysplasia
Authors: Slaughter Jonathan L. , Stenger Michael R. , Reagan Patricia B.
Published: March 2013

OBJECTIVES:

To determine (1) between-hospital variation in diuretic use for infants with bronchopulmonary dysplasia (BPD), including hospital-specific treatment frequency, treatment duration, and percentage of infants receiving short (≤5 consecutive days) versus longer (>5 days) courses, and to determine (2) demographic and clinical variables associated with diuretic administration.

METHODS:

A retrospective cohort study was conducted with the use of the Pediatric Health Information System to determine between-hospital variation in diuretic utilization patterns (primary outcome) and variables associated with diuretic use among <29-week-gestation infants with evolving BPD at age 28 days who were discharged between January 2007 and June 2011.

RESULTS:

During the 54-month study period, 1429 infants within 35 hospitals met the inclusion criteria for BPD at age 28 days, with 1222 (86%) receiving diuretic therapy for a median of 9 days (25th–75th percentile: 2–33 days). Short courses were administered to 1203 (83%) infants, and 570 (40%) infants received treatment for >5 consecutive days. Furosemide was the most widely prescribed diuretic (1218 infants; 85%), although chlorothiazide had the longest median duration of use (21 days; 25th–75th percentile: 8–46 days). The range of infants receiving a diuretic course of >5 days duration varied by hospital from 4% to 86%, with wide between-hospital variation even after adjustment for confounding variables.

CONCLUSIONS:

The frequency of diuretic administration to infants with BPD at US children’s hospitals, as well as the specific diuretic regimen used, varies markedly by institution. Safety and effectiveness research of long-term diuretic therapy for BPD patients is needed to develop evidence-based recommendations.
Efficacy of Psychosocial Group Intervention for Children With Chronic Illness and Their Parents
Authors: Scholten Linde , Willemen Agnes M. , Last Bob F. , Maurice-Stam Heleen , van Dijk Elisabeth M. , Ensink Elske , Zandbelt Noortje , van der Hoop-Mooij Aafke , Schuengel Carlo , Grootenhuis Martha A.
Published: March 2013

OBJECTIVE:

To investigate the efficacy of a cognitive-behavioral group intervention for children with chronic illnesses and to test the effect of an added parent component.

METHODS:

Children (n = 194) and their parents participated in a multicenter randomized clinical trial comparing a child-only intervention and a parent-child intervention to a wait-list control group. Primary outcomes were parent- and self-reported internalizing and externalizing problems; secondary outcomes were child disease-related coping skills (information seeking, relaxation, social competence, medical compliance, and positive thinking). Assessments took place at baseline and at 6- and 12-month follow-ups. Intention-to-treat mixed-model analyses were performed to test the difference in change in outcomes.

RESULTS:

The intervention had a positive effect on changes in parent-reported internalizing problems, child-reported externalizing problems, information seeking, social competence, and positive thinking. The additional effect of parental involvement was observed on parent-reported internalizing problems, child-reported externalizing problems, information seeking, and social competence. Illness severity and illness type did not moderate the effects. There were no intervention effects on child-reported internalizing problems, parent-reported externalizing problems, relaxation, or medical compliance. Of the families in the wait-list control group, 74% sought alternative psychological support during the intervention period.

CONCLUSIONS:

This RCT supports the efficacy of a protocol-based group intervention for children with chronic illnesses and their parents. Adding a parental component to the intervention contributed to the persistence of the effects. Future research should focus on moderating and mediating effects of the intervention.
Caustic Ingestion: A Possible Cause of Eosinophilic Esophagitis?
Authors: Homan Matjaž , Orel Rok , Liacouras Chris
Published: March 2013

Eosinophilic esophagitis (EoE) is an emerging disease in both pediatric and adult patients. It is a chronic disease of the esophagus and refers to intense eosinophilic infiltration limited to the esophageal epithelium in the absence of gastroesophageal reflux disease. In most patients, EoE is thought to be part of an allergic response to food antigens or aeroallergens. One such trigger could be caustic damage of the mucosa. To the best of our knowledge, the following case report describes for the first time the possible association between caustic injury of the esophagus and EoE.
On the use of formulations in person-centred, solution-focused short-term psychotherapy
Authors: Fitzgerald Pamela Elizabeth , Leudar Ivan
Published: March 2013

According to Carl Rogers, therapy must be nondirective in order to be effective. This means that the therapist needs to be trained to work within the clients’ frame of reference and do so in their practice. Conversation analytic research, however, suggests that therapists who claim to practise non-directive, non-authoritarian therapy nevertheless exercise subtle means of influencing their clients (e.g. through active listening, see Fitzgerald and Leudar 2010). The questions are: what in practice counts as being non-directive and how (relatively) nondirective psychotherapy is accomplished in practice. The present paper focuses on formulations which are one of the therapist's most useful tools and we demonstrate how these are used to guide clients to think along lines conducive to change.
Exploring lexical gaps in Australian Sign Language for the purposes of health communication
Authors: Major George , Napier Jemina , Ferrara Lindsay , Johnston Trevor
Published: March 2013

Until recently no linguistic research had been carried out on Australian Sign Language (Auslan) with regard to its use in healthcare settings, although anecdotal information suggests that the health lexicon of Auslan is strikingly under-developed. This paper describes a study that examined health terminology from the perspective of deaf people. Based on a thematic analysis of focus group data, the paper outlines strategies that deaf Australians report using when communicating about health concepts that do not have established signs in Auslan. Participants described a variety of communicative strategies – including the use of fingerspelling, depiction and explanations – that they employ in order to circumnavigate lexical gaps when talking about health. The study provides a crucial starting point for a discussion of the implications of lexical gaps in Auslan and highlights issues of particular relevance for interpreters who take responsibility for brokering health terms and mediating health communication.
The person in the room: How relating holistically contributes to an effective patient-care provider alliance
Authors: Penner Leslie A. , Roger Kerstin
Published: March 2013

The purpose of this paper is to explore how relating to the ‘whole’ person – both the physical body and the invisible aspects of the ‘self’ – is essential in the establishment of a strong therapeutic alliance between patients and health care providers. Our work is based on interviews conducted with individuals affected by neurological illnesses (patients and family care providers). Hsieh and Shannon's (2005) conventional content analysis was used to analyze the data. Under the broad theme of ‘maintaining a coherent sense of self’ we identified four distinct sub-themes related to interactions with health care providers. The results elucidate the more complex and deep needs of patients who must access care on an ongoing basis, and highlight the important role that care providers play in supporting individuals who are experiencing physical, spiritual and social losses. Care must attend to the deep needs of these individuals by communicating in a style that addresses both emotional and cognitive needs of patients, by thorough and holistic assessment and by appropriate referrals.
Tillering and forage accumulation in Marandu grass under different grazing intensities
Authors: Calvano Maria Paula Cavuto Abrão , Euclides Valéria Pacheco Batista , Montagner Denise Baptaglin , Lempp Beatriz , Difante Gelson dos Santos , Flores Renata Santos , Galbeiro Sandra
Published: March 2013

Studies of plant responses to defoliation are important to develop pasture management strategies. The objective of this study was to evaluate the population density of basal, aerial and reproductive tillers, tiller appearance and mortality rates, forage accumulation and sward structure in Marandu grass pastures under different grazing intensities. The experimental period was from January to June 2006, divided in three seasons: summer, autumn and winter. The pastures were continuously grazed using variable stocking rates. The grazing intensities corresponded to 15, 30 and 45 cm of sward height. The experiment was arranged in a complete randomized block design with three treatments and two replicates. The sward heights were measured twice a week. The response variables were: forage accumulation, forage mass and its morphological components; and population densities of basal (TPDb), aerial (TPDa) and reproductive (TPDr) tillers. The highest TPDb (P > 0.05) was recorded for the shortest sward pasture, and the highest TPDr (P<0.05) for the tallest sward pasture. Swards showed a tiller size/density compensation mechanism and, consequently, the forage accumulation was similar (P > 0.05) among the grazing intensities. Pasture with a sward height of 35 cm had 94% of sun light interception. The highest variations in forage accumulation and sward structure were more influenced by seasonal differences than by grazing intensities. Pastures of Marandu grass showed large flexibility in grazing management, which allowed it to be maintained at sward heights between 15 and 35 cm.