bluuebunny/crossref_metadata_2025 huggingface dataset:
Thoughts of suicide and stage of recovery in patients with schizophrenia in community mental health care
Authors: Gale Christopher , Skegg Keren , Mullen Richard , Patterson Tess , Gray Andrew
Published: July 2012

Objective:

Suicidality among patients with schizophrenia remains a major concern. The consumer-oriented concept of recovery has been recognised as important in recent years. We aimed to assess whether there was any relationship between stages of recovery and suicidality among people with schizophrenia in community care.

Method:

In a community mental health service, 85 patients with schizophrenia consented to an interview during which their stages of recovery were assessed using the Stages of Recovery Instrument. They were asked about suicidality and screened for depression using the Revised Clinical Interview Schedule. Insight was assessed via the Insight and Treatment Attitudes Questionnaire. Data analysis involved logistic regression.

Results:

In the past year 14% of the patients reported having wanted to die and 13% had considered suicide. Scores indicating the highest stage of recovery were associated with lower odds of suicidal thoughts. In contrast, suicidality was not significantly associated with depression or stage of insight.

Conclusion:

The link found between suicidality and stages of recovery has practical implications for suicide prevention among those with schizophrenia in community care. Approaches that facilitate the recovery process may reduce suicidal thoughts in this population.
Re-imaging: Mural painting and the state in Northern Ireland
Authors: Rolston Bill
Published: July 2012

This article focuses on the long history of mural painting in Northern Ireland and specifically on the changing relationship between mural painting and the state in various eras. Originally unionist murals were state-friendly, painted as part of the annual celebration of King William’s victory at the Battle of the Boyne in 1690. Later, murals were seen as beyond the pale, each for their own reasons; republican murals supported the armed struggle of the IRA against the state, while loyalist murals glorified the campaign of the loyalist paramilitary groups to terrorise the nationalist population. As the peace process took hold in Northern Ireland, republicans began to transform their own images; loyalists found this task more problematic, leading the state to intervene to fund the re-imaging of murals, thereby seeking to remove all offensive and military iconography. The article ends by critically assessing the outcome of the state’s Re-imaging Communities Programme.
Imagined intergroup contact promotes cooperation through outgroup trust
Authors: Pagotto Lisa , Visintin Emilio Paolo , De Iorio Giulia , Voci Alberto
Published: July 2012

The present research aims to investigate whether salience of memberships during imagined contact is necessary for producing generalized positive attitudes toward the outgroup and promoting intergroup cooperation. After a warm-up task that involved reciprocal self-disclosure during the imagined interaction with an outgroup member, we manipulated interpersonal versus intergroup features of imagined contact. Results indicated that participants who imagined a conversation with a Muslim focusing on intergroup differences subsequently reported more positive attitudes and cooperative intentions toward Muslim immigrants compared to either participants who imagined the interaction at the interpersonal level or participants in a control condition. Moreover, these effects were found to be mediated by outgroup trust. These findings attest to the strength of interventions based on imagined intergroup contact and suggest a possible implementation of the technique.
Zymographic differentiation of [NiFe]-Hydrogenases 1, 2 and 3 of Escherichia coli K-12
Authors: Pinske Constanze , Jaroschinsky Monique , Sargent Frank , Sawers Gary
Published: July 2012

Background

When grown under anaerobic conditions, Escherichia coli K-12 is able to synthesize three active [NiFe]-hydrogenases (Hyd1-3). Two of these hydrogenases are respiratory enzymes catalysing hydrogen oxidation, whereby Hyd-1 is oxygen-tolerant and Hyd-2 is considered a standard oxygen-sensitive hydrogenase. Hyd-3, together with formate dehydrogenase H (Fdh-H), forms the formate hydrogenlyase (FHL) complex, which is responsible for H2 evolution by intact cells. Hydrogen oxidation activity can be assayed for all three hydrogenases using benzyl viologen (BV; E
o′ = -360 mV) as an artificial electron acceptor; however ascribing activities to specific isoenzymes is not trivial. Previously, an in-gel assay could differentiate Hyd-1 and Hyd-2, while Hyd-3 had long been considered too unstable to be visualized on such native gels. This study identifies conditions allowing differentiation of all three enzymes using simple in-gel zymographic assays.

Results

Using a modified in-gel assay hydrogen-dependent BV reduction catalyzed by Hyd-3 has been described for the first time. High hydrogen concentrations facilitated visualization of Hyd-3 activity. The activity was membrane-associated and although not essential for visualization of Hyd-3, the activity was maximal in the presence of a functional Fdh-H enzyme. Furthermore, through the use of nitroblue tetrazolium (NBT; E
o′ = -80 mV) it was demonstrated that Hyd-1 reduces this redox dye in a hydrogen-dependent manner, while neither Hyd-2 nor Hyd-3 could couple hydrogen oxidation to NBT reduction. Hydrogen-dependent reduction of NBT was also catalysed by an oxygen-sensitive variant of Hyd-1 that had a supernumerary cysteine residue at position 19 of the small subunit substituted for glycine. This finding suggests that tolerance toward oxygen is not the main determinant that governs electron donation to more redox-positive electron acceptors such as NBT.

Conclusions

The utilization of particular electron acceptors at different hydrogen concentrations and redox potentials correlates with the known physiological functions of the respective hydrogenase. The ability to rapidly distinguish between oxygen-tolerant and standard [NiFe]-hydrogenases provides a facile new screen for the discovery of novel enzymes. A reliable assay for Hyd-3 will reinvigorate studies on the characterisation of the hydrogen-evolving FHL complex.
Two functional reticulocyte binding-like (RBL) invasion ligands of zoonotic Plasmodium knowlesi exhibit differential adhesion to monkey and human erythrocytes
Authors: Semenya Amma A , Tran Tuan M , Meyer Esmeralda VS , Barnwell John W , Galinski Mary R
Published: July 2012

Background

Plasmodium knowlesiis a monkey malaria species that is becoming a serious public health concern infecting hundreds and perhaps thousands of humans in Southeast Asia. Invasion of erythrocytes by merozoites entails a cascade of molecular interactions. One step involves the adhesion ofPlasmodiumreticulocyte binding-like (RBL) proteins.Plasmodium knowlesimerozoites express only two RBL invasion ligands, known as Normocyte Binding Proteins (PkNBPXa and PkNBPXb).

Methods

Overlapping N-terminal regions of PkNBPXa and PkNBPXb were expressed in COS7 cells and tested for surface expression and adhesion to rhesus monkey erythrocytes. Subsequent tests to study specific receptor ligand interactions included adhesion to a panel of human and non-human primate erythrocytes, enzymatic treatment, and site directed mutagenesis.

Results

An N-terminal cysteine-rich region of PkNBPXb (PkNBPXb-II) exhibited specific adhesion to rhesus monkey erythrocytes. Mutation of four of five cysteines in PkNBPXb-II interfered with its surface expression on COS7 cells, suggesting disulphide bond conformation is critical for intracellular trafficking. Binding of PkNBPXb-II was abolished when rhesus erythrocytes were pre-treated with chymotrypsin, but not trypsin or neuraminidase. PkNBPXb-II also bound other Old World monkey species and gibbon erythrocytes. However, erythrocytes from other primate species including humans did not bind to PkNBPXb-II or native PkNBPXb. Importantly, unlike PkNBPXb, PkNBPXa bound human erythrocytes, and this binding was independent of the Duffy blood group determinant.

Conclusions

The data reported here begins to clarify the functional domains of theP. knowlesiRBLs. A binding domain has been identified and characterized in PkNBPXb. Notably, this study demonstrates that unlike PkNBPXb, PkNBPXa can bind to human erythrocytes, suggesting that PkNBPXa may function as a ligand to enable the invasion ofP. knowlesimerozoites into human cells.
Characterization of grass carp reovirus minor core protein VP4
Authors: Yan Liming , Guo Hong , Sun Xiaoyun , Shao Ling , Fang Qin
Published: July 2012

Background

Grass Carp Reovirus (GCRV), a tentative member in the genus Aquareovirus of family Reoviridae, contains eleven segmented (double-stranded RNA) dsRNA genome which encodes 12 proteins. A low-copy core component protein VP4, encoded by the viral genome segment 5(S5), has been suggested to play a key role in viral genome transcription and replication.

Results

To understand the role of minor core protein VP4 played in molecular pathogenesis during GCRV infection, the recombinant GCRV VP4 gene was constructed and expressed in both prokaryotic and mammalian cells in this investigation. The recombinant His-tag fusion VP4 products expressed in E.coli were identified by Western blotting utilizing His-tag specific monoclonal and GCRV polyclonal antibodies. In addition, the expression of VP4 in GCRV infected cells, appeared in granules structure concentrated mainly in the cytoplasm, can be detected by Immunofluorescence (IF) using prepared anti-VP4 polyclonal antibody. Meanwhile, VP4 protein in GCRV core and infected cell lysate was identified by Immunoblotting (IB) assay. Of particular note, the VP4 protein was exhibited a diffuse distribution in the cytoplasm and nucleus in transfected cells, suggesting that VP4 protein may play a partial role in the nucleus by regulating cell cycle besides its predicted cytoplasmic function in GCRV infection.

Conclusions

Our results indicate the VP4 is a core component in GCRV. The cellular localization of VP4 is correlated with its predicted function. The data provide a foundation for further studies aimed at understanding the role of VP4 in viroplasmic inclusion bodies (VIB) formation during GCRV replication and assembly.
Peripheral T-cell Lymphoma with Cyclin D1 overexpression: a case report
Authors: Aquino Gabriella , Franco Renato , Ronconi Fioravante , Anniciello Annamaria , Russo Luigi , De Chiara Annarosaria , Panico Luigi
Published: July 2012

Peripheral T-cell lymphomas not otherwise specified are generally considered aggressive non-Hodgkin lymphomas, because of poor natural outcome and response to therapy. They show a complex karyotype without any specific genetic hallmark. We report a case of peripheral T-cell lymphoma not otherwise specified with heterogeneous nuclear Cyclin D1 immunohistochemical overexpression, due to gene copy gain, a phenomenon similar to that observed in Mantle Cell Lymphoma characterized by t(11;14)(q13;q32). In this case report we underline the diagnostic pitfall rapresented by Cyclin D1 immunoistochemical overexpression in a T-cell lymphoma. Several pitfalls could lead to misinterpretation of diagnosis, therefore, we underlined the need to integrate the classical histology and immunohistochemistry with molecular tests as clonality or Fluorescence in situ hybridization.

Virtual slide

The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1117747619703769
IL-6 regulation on skeletal muscle mitochondrial remodeling during cancer cachexia in the Apc
Min/+
mouse
Authors: White James P , Puppa Melissa J , Sato Shuichi , Gao Song , Price Robert L , Baynes John W , Kostek Matthew C , Matesic Lydia E , Carson James A
Published: July 2012

Background

Muscle protein turnover regulation during cancer cachexia is being rapidly defined, and skeletal muscle mitochondria function appears coupled to processes regulating muscle wasting. Skeletal muscle oxidative capacity and the expression of proteins regulating mitochondrial biogenesis and dynamics are disrupted in severely cachectic Apc

Min/+
mice. It has not been determined if these changes occur at the onset of cachexia and are necessary for the progression of muscle wasting. Exercise and anti-cytokine therapies have proven effective in preventing cachexia development in tumor bearing mice, while their effect on mitochondrial content, biogenesis and dynamics is not well understood. The purposes of this study were to 1) determine IL-6 regulation on mitochondrial remodeling/dysfunction during the progression of cancer cachexia and 2) to determine if exercise training can attenuate mitochondrial dysfunction and the induction of proteolytic pathways during IL-6 induced cancer cachexia.

Methods

Apc

Min/+
mice were examined during the progression of cachexia, after systemic interleukin (IL)-6r antibody treatment, or after IL-6 over-expression with or without exercise. Direct effects of IL-6 on mitochondrial remodeling were examined in cultured C2C12 myoblasts.

Results

Mitochondrial content was not reduced during the initial development of cachexia, while muscle PGC-1α and fusion (Mfn1, Mfn2) protein expression was repressed. With progressive weight loss mitochondrial content decreased, PGC-1α and fusion proteins were further suppressed, and fission protein (FIS1) was induced. IL-6 receptor antibody administration after the onset of cachexia improved mitochondrial content, PGC-1α, Mfn1/Mfn2 and FIS1 protein expression. IL-6 over-expression in pre-cachectic mice accelerated body weight loss and muscle wasting, without reducing mitochondrial content, while PGC-1α and Mfn1/Mfn2 protein expression was suppressed and FIS1 protein expression induced. Exercise normalized these IL-6 induced effects. C2C12 myotubes administered IL-6 had increased FIS1 protein expression, increased oxidative stress, and reduced PGC-1α gene expression without altered mitochondrial protein expression.

Conclusions

Altered expression of proteins regulating mitochondrial biogenesis and fusion are early events in the initiation of cachexia regulated by IL-6, which precede the loss of muscle mitochondrial content. Furthermore, IL-6 induced mitochondrial remodeling and proteolysis can be rescued with moderate exercise training even in the presence of high circulating IL-6 levels.
B lymphocyte-typing for prediction of clinical response to rituximab
Authors: Brezinschek Hans-Peter , Rainer Franz , Brickmann Kerstin , Graninger Winfried B
Published: July 2012

Introduction

The prediction of therapeutic response to rituximab in rheumatoid arthritis is desirable. We evaluated whether analysis of B lymphocyte subsets by flow cytometry would be useful to identify non-responders to rituximab ahead of time.

Methods

Fifty-two patients with active rheumatoid arthritis despite therapy with TNF-inhibitors were included in the national rituximab registry. DAS28 was determined before and 24 weeks after rituximab application. B cell subsets were analyzed by high-sensitive flow cytometry before and 2 weeks after rituximab administration. Complete depletion of B cells was defined as CD19-values below 0.0001 x109 cells/liter.

Results

At 6 months 19 patients had a good (37%), 23 a moderate (44%) and 10 (19%) had no EULAR-response. The extent of B lymphocyte depletion in peripheral blood did not predict the success of rituximab therapy. Incomplete depletion was found at almost the same frequency in EULAR responders and non-responders. In comparison to healthy controls, non-responders had elevated baseline CD95+ pre-switch B cells, whereas responders had a lower frequency of plasmablasts.

Conclusions

The baseline enumeration of B lymphocyte subsets is still of limited clinical value for the prediction of response to anti-CD20 therapy. However, differences at the level of CD95+ pre switch B cells or plasmablasts were noticed with regard to treatment response. The criterion of complete depletion of peripheral B cells after rituximab administration did not predict the success of this therapy in rheumatoid arthritis.
Survival and neurologic outcome after traumatic out-of-hospital cardiopulmonary arrest in a pediatric and adult population: a systematic review
Authors: Zwingmann Jörn , Mehlhorn Alexander T , Hammer Thorsten , Bayer Jörg , Südkamp Norbert P , Strohm Peter C
Published: July 2012

Introduction

This systematic review is focused on the in-hospital mortality and neurological outcome of survivors after prehospital resuscitation following trauma. Data were analyzed for adults/pediatric patients and for blunt/penetrating trauma.

Methods

A systematic review was performed using the data available in Ovid Medline. 476 articles from 1/1964 - 5/2011 were identified by two independent investigators and 47 studies fulfilled the requirements (admission to hospital after prehospital resuscitation following trauma). Neurological outcome was evaluated using the Glasgow outcome scale.

Results

34 studies/5391 patients with a potentially mixed population (no information was found in most studies if and how many children were included) and 13 paediatric studies/1243 children (age ≤ 18 years) were investigated. The overall mortality was 92.8% (mixed population: 238 survivors, lethality 96.7%; paediatric group: 237 survivors, lethality 86.4% = p < 0.001).

Penetrating trauma was found in 19 studies/1891 patients in the mixed population (69 survivors, lethality: 96.4%) and in 3 pediatric studies/91 children (2 survivors lethality 97.8%).

44.3% of the survivors in the mixed population and 38.3% in the group of children had a good neurological recovery. A moderate disability could be evaluated in 13.1% in the mixed population and in 12.8% in children. A severe disability was found in 29.5% of the survivors in the mixed patients and in 38.3% in the group of children. A persistent vegetative state was the neurological status in 9.8% in the mixed population and in 10.6% in children.

For each year prior to 2010, the estimated log-odds for survival decreased by 0.022 (95%-CI: [0.038;0.006]). When jointly analyzing the studies on adults and children, the proportion of survivors for children is estimated to be 17.8% (95%-CI: [15.1%;20.8%]). The difference of the paediatric compared to the adult proportion is significant (p < 0.001).

Conclusions

Children have a higher chance of survival after resuscitation of an out-of-hospital traumatic cardiac arrest compared to adults but tend to have a poorer neurological outcome at discharge.
Adrenal Steroidogenesis after B Lymphocyte Depletion Therapy in New-Onset Addison's Disease
Authors: Pearce Simon H. S. , Mitchell Anna L. , Bennett Stuart , King Phil , Chandran Sukesh , Nag Sath , Chen Shu , Smith Bernard Rees , Isaacs John D. , Vaidya Bijay
Published: July 2012

Context:

A diagnosis of Addison's disease means lifelong dependence on daily glucocorticoid and mineralocorticoid therapy and is associated with increased morbidity and mortality as well as a risk of unexpected adrenal crisis.

Objective:

The objective of the study was to determine whether immunomodulatory therapy at an early stage of autoimmune Addison's disease could lead to preservation or improvement in adrenal steroidogenesis.

Design and Intervention:

This was an open-label, pilot study of B lymphocyte depletion therapy in new-onset idiopathic primary adrenal failure. Doses of iv rituximab (1 g) were given on d 1 and 15, after pretreatment with 125 mg iv methylprednisolone.

Patients and Main Outcome Measures:

Six patients (aged 17–47 yr; four females) were treated within 4 wk of the first diagnosis of idiopathic primary adrenal failure. Dynamic testing of adrenal function was performed every 3 months for at least 12 months.

Results:

Serum cortisol levels declined rapidly and were less than 100 nmol/liter (3.6 μg/dl) in all patients by 3 months after B lymphocyte depletion. Serum cortisol and aldosterone concentrations remained low in five of the six patients throughout the follow-up period. However, a single patient had sustained improvement in both serum cortisol [peak 434 nmol/liter (15.7 μg/dl)] and aldosterone [peak 434 pmol/liter (15.7 ng/dl)] secretion. This patient was able to discontinue steroid medications 15 months after therapy and remains well, with improving serum cortisol levels 27 months after therapy.

Conclusion:

New-onset autoimmune Addison's disease should be considered as a potentially reversible condition in some patients. Future studies of immunomodulation in autoimmune Addison's disease may be warranted.
Simulations demonstrate a simple network to be sufficient to control branch point selection, smooth muscle and vasculature formation during lung branching morphogenesis
Authors: Cellière Géraldine , Menshykau Denis , Iber Dagmar
Published: July 2012

Proper lung functioning requires not only a correct structure of the conducting airway tree, but also the simultaneous development of smooth muscles and vasculature. Lung branching morphogenesis is strongly stereotyped and involves the recursive use of only three modes of branching. We have previously shown that the experimentally described interactions between Fibroblast growth factor (FGF)10, Sonic hedgehog (SHH) and Patched (Ptc) can give rise to a Turing mechanism that not only reproduces the experimentally observed wildtype branching pattern but also, in part counterintuitive, patterns in mutant mice. Here we show that, even though many proteins affect smooth muscle formation and the expression of Vegfa, an inducer of blood vessel formation, it is sufficient to add FGF9 to the FGF10/SHH/Ptc module to successfully predict simultaneously the emergence of smooth muscles in the clefts between growing lung buds, and Vegfa expression in the distal sub-epithelial mesenchyme. Our model reproduces the phenotype of both wildtype and relevant mutant mice, as well as the results of most culture conditions described in the literature.
The overexpression of nuclear envelope protein Lap2β induces endoplasmic reticulum reorganisation via membrane stacking
Authors: Volkova Ekaterina G. , Abramchuk Sergey S. , Sheval Eugene V.
Published: July 2012

Some nuclear envelope proteins are localised to both the nuclear envelope and the endoplasmic reticulum; therefore, it seems plausible that even small amounts of these proteins can influence the organisation of the endoplasmic reticulum. A simple method to study the possible effects of nuclear envelope proteins on endoplasmic reticulum organisation is to analyze nuclear envelope protein overexpression. Here, we demonstrate that Lap2β overexpression can induce the formation of cytoplasmic vesicular structures derived from endoplasmic reticulum membranes. Correlative light and electron microscopy demonstrated that these vesicular structures were composed of a series of closely apposed membranes that were frequently arranged in a circular fashion. Although stacked endoplasmic reticulum cisternae were highly ordered, Lap2β could readily diffuse into and out of these structures into the surrounding reticulum. It appears that low-affinity interactions between cytoplasmic domains of Lap2β can reorganise reticular endoplasmic reticulum into stacked cisternae. Although the effect of one protein may be insignificant at low concentrations, the cumulative effect of many non-specialised proteins may be significant.
Yolk syncytial layer formation is a failure of cytokinesis mediated by Rock1 function in the early zebrafish embryo
Authors: Chu Lee-Thean , Fong Steven H. , Kondrychyn Igor , Loh Siau Lin , Ye Zhanrui , Korzh Vladimir
Published: July 2012

The yolk syncytial layer (YSL) performs multiple critical roles during zebrafish development. However, little is known about the cellular and molecular mechanisms that underlie the formation of this important extraembryonic structure. Here, we demonstrate by timelapse confocal microscopy of a transgenic line expressing membrane-targeted GFP that the YSL forms as a result of the absence of cytokinesis between daughter nuclei at the tenth mitotic division and the regression of pre-existing marginal cell membranes, thus converting the former margin of the blastoderm into a syncytium. We show that disruption of components of the cytoskeleton induces the formation of an expanded YSL, and identify Rock1 as the regulator of cytoskeletal dynamics that lead to YSL formation. Our results suggest that the YSL forms as a result of controlled cytokinesis failure in the marginal blastomeres, and Rock1 function is necessary for this process to occur. Uncovering the cellular and molecular mechanisms underlying zebrafish YSL formation offers significant insight into syncytial development in other tissues as well as in pathological conditions.
A growth factor signaling cascade confined to circular ruffles in macrophages
Authors: Welliver Timothy P. , Swanson Joel A.
Published: July 2012

The formation of macropinosomes requires large-scale movements of membranes and the actin cytoskeleton. Over several minutes, actin-rich surface ruffles transform into 1–5 µm diameter circular ruffles, which close at their distal margins, creating endocytic vesicles. Previous studies using fluorescent reporters of phosphoinositides and Rho-family GTPases showed that signals generated by macrophages in response to the growth factor Macrophage Colony-Stimulating Factor (M-CSF) appeared transiently in domains of plasma membrane circumscribed by circular ruffles. To address the question of how signaling molecules are coordinated in such large domains of plasma membrane, this study analyzed the relative timing of growth factor-dependent signals as ruffles transformed into macropinosomes. Fluorescent protein chimeras expressed in macrophages were imaged by microscopy and quantified relative to circular ruffle formation and cup closure. The large size of macropinocytic cups allowed temporal resolution of the transitions in phosphoinositides and associated enzyme activities that organize cup closure. Circular ruffles contained transient and sequential spikes of phosphatidylinositol (4,5)-bisphosphate (PI(4,5)P2), phosphatidylinositol (3,4,5)-trisphosphate (PIP3), diacylglycerol, PI(3,4)P2, PI(3)P and the activities of protein kinase C-α, Rac1, Ras and Rab5. The confinement of this signal cascade to circular ruffles indicated that diffusion barriers present in these transient structures focus feedback activation and deactivation of essential enzyme activities into restricted domains of plasma membrane.
Fss/Tbx6 is required for central dermomyotome cell fate in zebrafish
Authors: Windner Stefanie Elisabeth , Bird Nathan Craig , Patterson Sara Elizabeth , Doris Rosemarie Anne , Devoto Stephen Henri
Published: July 2012

The dermomyotome is a pool of progenitor cells on the surface of the myotome. In zebrafish, dermomyotome precursors (anterior border cells, ABCs) can be first identified in the anterior portion of recently formed somites. They must be prevented from undergoing terminal differentiation during segmentation, even while mesodermal cells around them respond to signaling cues and differentiate.

T-box containing transcription factors regulate many aspects of mesoderm fate including segmentation and somite patterning. The fused somites (fss) gene is the zebrafish ortholog of tbx6. We demonstrate that in addition to its requirement for segmentation, fss/tbx6 is also required for the specification of ABCs and subsequently the central dermomyotome. The absence of Tbx6-dependent central dermomyotome cells in fss/tbx6 mutants is spatially coincident with a patterning defect in the myotome.

Using transgenic fish with a heat-shock inducible tbx6 gene in the fss/tbx6 mutant background, we further demonstrate that ubiquitous fss/tbx6 expression has spatially distinct effects on recovery of the dermomyotome and segment boundaries, suggesting that the mechanism of Fss/Tbx6 action is distinct with respect to dermomyotome development and segmentation.

We propose that Fss/Tbx6 is required for preventing myogenic differentiation of central dermomyotome precursors before and after segmentation and that central dermomyotome cells represent a genetically and functionally distinct subpopulation within the zebrafish dermomyotome.
Regulation of behavioral circadian rhythms and clock protein PER1 by the deubiquitinating enzyme USP2
Authors: Yang Yaoming , Duguay David , Bédard Nathalie , Rachalski Adeline , Baquiran Gerardo , Na Chan Hyun , Fahrenkrug Jan , Storch Kai-Florian , Peng Junmin , Wing Simon S. , Cermakian Nicolas
Published: July 2012

Endogenous 24-hour rhythms are generated by circadian clocks located in most tissues. The molecular clock mechanism is based on feedback loops involving clock genes and their protein products. Post-translational modifications, including ubiquitination, are important for regulating the clock feedback mechanism. Previous work has focused on the role of ubiquitin ligases in the clock mechanism. Here we show a role for the rhythmically-expressed deubiquitinating enzyme ubiquitin specific peptidase 2 (USP2) in clock function. Mice with a deletion of the Usp2 gene (Usp2 KO) display a longer free-running period of locomotor activity rhythms and altered responses of the clock to light. This was associated with altered expression of clock genes in synchronized Usp2 KO mouse embryonic fibroblasts and increased levels of clock protein PERIOD1 (PER1). USP2 can be coimmunoprecipitated with several clock proteins but directly interacts specifically with PER1 and deubiquitinates it. Interestingly, this deubiquitination does not alter PER1 stability. Taken together, our results identify USP2 as a new core component of the clock machinery and demonstrate a role for deubiquitination in the regulation of the circadian clock, both at the level of the core pacemaker and its response to external cues.
Accelerated renal disease is associated with the development of metabolic syndrome in a glucolipotoxic mouse model
Authors: Martínez-García Cristina , Izquierdo Adriana , Velagapudi Vidya , Vivas Yurena , Velasco Ismael , Campbell Mark , Burling Keith , Cava Fernando , Ros Manuel , Orešič Matej , Vidal-Puig Antonio , Medina-Gomez Gema
Published: July 2012

Individuals with metabolic syndrome are at high risk of developing chronic kidney disease (CKD) through unclear pathogenic mechanisms. Obesity and diabetes are known to induce glucolipotoxic effects in metabolically relevant organs. However, the pathogenic role of glucolipotoxicity in the aetiology of diabetic nephropathy is debated. We generated a murine model, the POKO mouse, obtained by crossing the peroxisome proliferator-activated receptor gamma 2 (PPARγ2) knockout (KO) mouse into a genetically obese ob/ob background. We have previously shown that the POKO mice showed: hyperphagia, insulin resistance, hyperglycaemia and dyslipidaemia as early as 4 weeks of age, and developed a complete loss of normal β-cell function by 16 weeks of age. Metabolic phenotyping of the POKO model has led to investigation of the structural and functional changes in the kidney and changes in blood pressure in these mice. Here we demonstrate that the POKO mouse is a model of renal disease that is accelerated by high levels of glucose and lipid accumulation. Similar to ob/ob mice, at 4 weeks of age these animals exhibited glomerulomegaly, an increased urinary albumin:creatinine ratio and significantly increased blood pressure, but in contrast showed a significant increase in the renal hypertrophy index and an associated increase in p27Kip1 expression compared with their obese littermates. Moreover, at 4 weeks of age POKO mice showed insulin resistance, an alteration of lipid metabolism and glomeruli damage associated with significantly increased transforming growth factor beta (TGFβ) and parathyroid hormone-related protein (PTHrP) expression. At this age, levels of proinflammatory molecules, such as monocyte chemoattractant protein-1 (MCP-1), and fibrotic factors were also increased at the glomerular level compared with levels in ob/ob mice. At 12 weeks of age, renal damage was fully established. These data suggest an accelerated lesion through glucolipotoxic effects in the renal pathogenesis in POKO mice.
Facial dysmorphism induced by bone morphogenetic protein receptor type 1A-mediated signaling reduction in neural crest-derived cells
Authors: Saito Hiromitsu , Yamamura Ken-ichi , Suzuki Noboru
Published: July 2012

Bone morphogenetic protein (BMP) receptor type 1A (BMPR1A) mutations are associated with facial dysmorphism, which is one of the main clinical signs in both juvenile polyposis and chromosome 10q23 deletion syndromes. Craniofacial development requires reciprocal epithelial/neural crest (NC)-derived mesenchymal interactions mediated by signaling factors, such as Bmp, in both cell populations. To address the role of mesenchymal Bmp signaling in craniofacial development, we generated a conditional knockdown mouse by expressing the dominant-negative Bmpr1a in NC-derived cells expressing the Myelin protein zero (Mpz)-Cre transgene. At birth, 100% of the conditional mutant mice had wide-open anterior fontanelles, and 80% of them died because of cleft face and cleft palate soon after birth. The other 20% survived and developed short faces, hypertelorism, and calvarial foramina. Analysis of the NC-derived craniofacial mesenchyme of mutant embryos revealed an activation of P53 apoptosis pathway, and down-regulation of both c-Myc and Bcl-XL; a normal growth rate but an incomplete expansion of mesenchymal cells. These findings provide genetic evidence indicating that optimal Bmpr1a-mediated signaling is essential for NC-derived mesenchymal cell survival in both normal nasal and frontal bone development and suggest that our model is useful for studying some aspects of the molecular etiology of human craniofacial dysmorphism.
Zebrafish neurofibromatosis type 1 genes have redundant functions in tumorigenesis and embryonic development
Authors: Shin Jimann , Padmanabhan Arun , de Groh Eric D. , Lee Jeong-Soo , Haidar Sam , Dahlberg Suzanne , Guo Feng , He Shuning , Wolman Marc A. , Granato Michael , Lawson Nathan D. , Wolfe Scot A. , Kim Seok-Hyung , Solnica-Krezel Lilianna , Kanki John P. , Ligon Keith L. , Epstein Jonathan A. , Look A. Thomas
Published: July 2012

Neurofibromatosis type 1 (NF1) is a common, dominantly inherited genetic disorder that results from mutations in the neurofibromin 1 (NF1) gene. Affected individuals demonstrate abnormalities in neural crest-derived tissues that include hyperpigmented skin lesions and benign peripheral nerve sheath tumors. NF1 patients also have a predisposition to malignancies including juvenile myelomonocytic leukemia (JMML), optic glioma, glioblastoma, schwannoma, and malignant peripheral nerve sheath tumors (MPNSTs). In an effort to better define the molecular and cellular determinants of NF1 disease pathogenesis in vivo, we employed targeted mutagenesis strategies to generate zebrafish harboring stable germline mutations in nf1a and nf1b, orthologues of NF1. Animals homozygous for loss-of-function alleles of nf1a or nf1b alone are phenotypically normal and viable. Homozygous loss of both alleles in combination generates larval phenotypes that resemble aspects of the human disease and results in larval lethality between 7 and 10 days post fertilization. nf1-null larvae demonstrate significant central and peripheral nervous system defects. These include aberrant proliferation and differentiation of oligodendrocyte progenitor cells (OPCs), dysmorphic myelin sheaths, and hyperplasia of Schwann cells. Loss of nf1 contributes to tumorigenesis as demonstrated by an accelerated onset and increased penetrance of high-grade gliomas and MPNSTs in adult nf1a+/-; nf1b-/-; p53e7/e7 animals. nf1-null larvae also demonstrate significant motor and learning defects. Importantly, we identify and quantitatively analyze a novel melanophore phenotype in nf1-null larvae, providing the first animal model of the pathognomonic pigmentation lesions of NF1. Together, these findings support a role for nf1a and nf1b as potent tumor suppressor genes that also function in the development of both central and peripheral glial cells as well as melanophores in zebrafish.
Oxidative stress contributes to outcome severity in a Drosophila melanogaster model of classic galactosemia
Authors: Jumbo-Lucioni Patricia P. , Hopson Marquise L. , Hang Darwin , Liang Yongliang , Jones Dean P. , Fridovich-Keil Judith L.
Published: July 2012

Classic galactosemia is a genetic disorder that results from profound loss of galactose-1P-uridylyltransferase (GALT). Affected infants experience a rapid escalation of potentially lethal acute symptoms following exposure to milk. Dietary restriction of galactose prevents or resolves the acute sequelae; however, many patients experience profound long-term complications. Despite decades of research, the mechanisms that underlie pathophysiology in classic galactosemia remain unclear. Recently, we developed a Drosophila melanogaster model of classic galactosemia and demonstrated that, like patients, GALT-null Drosophila succumb in development if exposed to galactose but live if maintained on a galactose-restricted diet. Prior models of experimental galactosemia have implicated a possible association between galactose exposure and oxidative stress. Here we describe application of our fly genetic model of galactosemia to the question of whether oxidative stress contributes to the acute galactose sensitivity of GALT-null animals. Our first approach tested the impact of pro- and antioxidant food supplements on the survival of GALT-null and control larvae. We observed a clear pattern: the oxidants paraquat and DMSO each had a negative impact on the survival of mutant but not control animals exposed to galactose, and the antioxidants vitamin C and α-mangostin each had the opposite effect. Biochemical markers also confirmed that galactose and paraquat synergistically increased oxidative stress on all cohorts tested but, interestingly, the mutant animals showed a decreased response relative to controls. Finally, we tested the expression levels of two transcripts responsive to oxidative stress, GSTD6 and GSTE7, in mutant and control larvae exposed to galactose and found that both genes were induced, one by more than 40-fold. Combined, these results implicate oxidative stress and response as contributing factors in the acute galactose sensitivity of GALT-null Drosophila and, by extension, suggest that reactive oxygen species might also contribute to the acute pathophysiology in classic galactosemia.
Schwann Cell Myelination Requires Integration of Laminin Activities
Authors: McKee Karen K. , Yang Dong-Hua , Patel Rajesh , Chen Zu-Lin , Strickland Sidney , Takagi Junichi , Sekiguchi Kiyotoshi , Yurchenco Peter D.
Published: July 2012

Laminins promote early stages of peripheral nerve myelination by assembling basement membranes (BMs) on Schwann cell surfaces, leading to activation of β1-integrins and other receptors. The BM composition, structural bonds and ligands needed to mediate this process, however, are not well understood. A mouse hypomorphic for laminin γ1-subunit expression that assembled endoneurial BMs with reduced component density exhibited an axonal sorting defect with amyelination but normal Schwann cell proliferation, the latter unlike the null. To identify the basis for this, and to dissect participating laminin interactions, Lamc1 gene-inactivated dorsal root ganglia were treated with recombinant laminins-211 and -111 lacking different architecture-forming and receptor-binding activities to induce myelination. Myelin-wrapping of axons by Schwann cells was found to require higher laminin concentrations than either proliferation or axonal ensheathment. Laminins that were unable to polymerize through deletions that removed critical LN-domains, or that lacked cell-adhesive LG-domains, caused reduced BMs and nearly absent myelination. Laminins engineered to bind weakly to α6β1 and/or α7β1 integrins through their LG-domains, even though they could effectively assemble BMs, decreased myelination. Proliferation depended upon both integrin-binding to LG domains and polymerization. Collectively these findings reveal that laminins integrate scaffold-forming and cell-adhesion activities to assemble an endoneurial BM, with myelination and proliferation requiring additional α6β1/α7β1-laminin LG-domain interactions, and that a high BM ligand/structural-density is needed for efficient myelination.
Human mesenchymal stem cells shift CD8+ T cells towards a suppressive phenotype by inducing tolerogenic monocytes
Authors: Hof-Nahor Irit , Leshansky Lucy , Shivtiel Shoham , Eldor Liron , Aberdam Daniel , Itskovitz-Eldor Joseph , Berrih-Aknin Sonia
Published: July 2012

The mechanisms underlying the immunomodulatory effects of mesenchymal stem cells (MSCs) have been essentially studied in conditions of strong T cell activation that represents extreme situation and induces rapid death of activated lymphocytes. The objective of this study was to investigate these mechanisms in absence of additional polyclonal activation. In cocultures of peripheral mononuclear blood cells with hMSC, we observed a striking decreased expression of CD8 level on CD8+ cells, together with decreased CD28 and CD44 expression and impaired IFN-gamma and Granzyme B production. This effect was specific to hMSCs, since it was not observed with several other cell lines. Down-regulation of CD8 expression required CD14+ monocytes in direct contact with the CD8+ cells, while the effects of hMSCs on the CD14+ cells were essentially mediated by soluble factors. The CD14+ monocytes exhibited a tolerogenic pattern when co-cultured with hMSCs, with a clear decrease in CD80 and CD86 co-stimulatory molecules, and an increase in the inhibitory receptors ILT-3 and ILT-4. MSC-preconditioned CD8+ cells had similar effects on monocytes and were able to inhibit lymphocyte proliferation. Injection of human MSCs in humanized NSG mice showed similar trends, in particular decreased CD44 and CD28 on human immune cells. Altogether, our study demonstrates a new immunomodulation mechanism of action of hMSCs through the modulation of CD8+ cells towards a non-cytotoxic/suppressive phenotype. This mechanism of action has to be taken into account in clinical trials, where it should be beneficial in grafts and autoimmune diseases, but potentially detrimental in malignant diseases.
Retention Behavior of β-Cyclodextrin-bonded Stationary Phase Synthesized via Atom-transfer Radical Polymerization and Click Chemistry
Authors: Wang Huaisong , Xie Qiwei , Wang Haiyan , Zhu Diankui , Jiang An
Published: July 2012

A new chiral stationary phase was designed and synthesized via surface-initiated atom-transfer radical polymerization (ATRP) and click chemistry. Poly(2-methyl-3-butyn-2-yl methacrylate-co-divinylbenzene), poly(MBMA-co-DVB), was grafted on silica surface via ATRP (“grafting-from” technique) first, and then azide-modified β-CD was immobilized on the alkyne of the polymer layer as chiral separation material. Different crosslinking degree of poly(MBMA-co-DVB) was selected to improve the hydrophobicity of silica surface. The materials were packed into a stainless-steel column (150 mm × 4.6 mm i.d.) for high-performance liquid chromatography (HPLC), and evaluated by separation of aromatic and chiral compounds. The result demonstrated that the crosslinked benzene ring in poly(MBMA-co-DVB) can well improve the selectivity and retention time.
Involving interpreters in research studies
Authors: Plumridge Gillian , Redwood Sabi , Greenfield Sheila , Akhter Nasreen , Chowdhury Raja , Khalade Abdul , Gill Paramjit
Published: July 2012

It is important to include non English speakers in health services research to ensure not only the generalisability of findings but also to address health inequalities and promote social justice. One approach is to use interpreters but there is little guidance for working with an interpreter in interviews. Involving an interpreter presents challenges in the planning and conduct of research interviews which can be minimised by an awareness of various theoretical and practical implications and of potential pitfalls. Drawing both on our experiences of involving interpreters in research and on the literature, we raise some issues that researchers need to consider.
Grading of structural timber of Populus × euramericana clone I-214
Authors: Casado Milagros , Acuña Luis , Basterra Luis-Alfonso , Ramón-Cueto Gemma , Vecilla Daniel
Published: July 2012

Grading tests were performed on 338 poplar wood samples (Populus × euramericana I-214) with dimensions of 150×50 mm2, 150×80 mm2, and 200×100 mm2 prepared for structural utilization. The non-destructive testing methods included ultrasound (Sylvatest) and induced vibrations (Portable Lumber Grader) combined with visual grading criteria. Additionally, screw withdrawal resistance was tested by the instrument Fakopp. These results were compared with those obtained by a four-point bending test according to the EN 408 (2004) standard. Ultrasound and induced vibrations overestimated the quality of this type of wood. Correlation equations are proposed to determine the true characteristics for each testing method. It can be concluded that poplar timber from clone I-214 has acceptable flexural strength, but a comparatively low elastic modulus, whereby it is not graded within a high strength class according to the EN 338 (2009) standard.
Isolation and characterization of β-cadinene synthase cDNA from Chamaecyparis formosensis Matsum
Authors: Kuo Pei-Min , Hsu Keng-Hao , Lee Yi-Ru , Chu Fang-Hua , Wang Sheng-Yang
Published: July 2012

Chamaecyparis formosensis is a precious conifer endemic in Taiwan. To understand the sesquiterpene synthesis mechanism in this tree, full-length cDNA of a putative sesquiterpene synthase (sesqui-TPS), designated Cf-Cad, was obtained by rapid amplification of complementary DNA ends-polymerase chain reaction. Cf-Cad is 1812 bp in length. To identify its function, recombinant protein from Escherichia coli was incubated with farnesyl diphosphate, which produced one major product, the structure of which was elucidated by gas chromatography/mass spectrometry (GC/MS) analysis. GC/MS analysis, GC retention time and MS matching with authentic standards revealed that the major product was β-cadinene. This is the first report of the cloning, functional expression in E. coli and identification of a sesqui-TPS from a Cupressaceae conifer.
Fundamentals of Biobased and Biodegradable Lubricants: A Real World Perspective
Authors: Miller Mark
Published: July 2012

Environmentally friendly lubricants, which are biodegradable and non-toxic, are essential in industrial applications. Significant quantities of petroleum fluids enter the environment every year through leaks, spills, line breakage, and fitting failures. The green alternative helps to decrease environmental impact, as well as clean up costs and fines.