bluuebunny/crossref_metadata_2025 huggingface dataset:
Framing sustainability: A case study analysis of the environment and sustainability discourse in the Indian English language press
Authors: Nambiar Prithi
Published: January 2014

The definition and local cultural adoption of globally generated terms such as environment and sustainable development happen largely through an imbricated process of meaning negotiation within democratic public discourse. Framing behaviour by media and influential opinion makers shapes and moulds local and national discourse about environment and sustainability, particularly in developing countries where pro-development forces face off against pro-conservation groups over resource use choices. Using elements of framing and grounded theory, a qualitative interpretive case study analysis of environment and sustainability related articles from a leading English language fortnightly magazine in India reveals that highly emotive frames are being used in media construction of the environment and sustainability discourse in India. While the research findings suggest that stock themes from the international framing repertoire are in use, they also indicate that distinctive new frames are being employed that could assist in instigating social change and refashioning public attitudes.
Noise-induced hearing loss
Authors: Nair Chitra
Published: January 2014

Noise-induced hearing loss is the permanent hearing impairment that results from prolonged exposure to high levels of noise. Hearing loss caused by recreational and occupational noise exposure is the second-most-common cause of sensorineural hearing loss after presbycusis. Noise exposure is a potentially avoidable cause of deafness and the World Health Organisation recommends integrating elements of prevention, diagnosis, education, hearing promotion and conservation into primary care.
Endovascular Therapy for Acute Stroke in Patients With Cancer
Authors: Merkler Alexander E. , Marcus Justin R. , Gupta Ajay , Kishore Sirish A. , Leifer Dana , Patsalides Athos , DeAngelis Lisa M. , Navi Babak B.
Published: January 2014

Intravenous thrombolysis is the standard treatment for acute ischemic stroke (AIS). However, patients with cancer who have stroke are often precluded from therapy because of coagulopathy or recent surgery. Endovascular therapy may be a more suitable recanalization strategy for some patients with cancer and stroke, but no prior detailed reports documenting its use in this population exist. We present a case series from a tertiary care referral center of 2 patients with active systemic cancer who were successfully treated with endovascular therapy for AIS. Both patients had active lung cancer with excellent premorbid functional status and presented with severe AIS from left middle cerebral artery occlusions. Intravenous thrombolysis was deferred because of absolute contraindications. Mechanical embolectomy was performed instead and revascularization was achieved within 5 hours in both patients, resulting in dramatic neurological recoveries— National Institutes of Health Stroke Scale improved from 14 to 0 and from 23 to 3 from admission to discharge, respectively. In conclusion, endovascular therapy may be beneficial for select patients with cancer and AIS who are ineligible for intravenous thrombolysis. However, further studies are needed to determine the safety and efficacy of endovascular therapy in the population with cancer.
Late Onset of CSF Rhinorrhea in a Postoperative Transsphenoidal Surgery Patient Following Robotic-Assisted Abdominal Hysterectomy
Authors: Dowdy Justin T. , Moody Marcus W. , Cifarelli Christopher P.
Published: January 2014

Cerebrospinal fluid (CSF) leak is the most commonly encountered perioperative complication in transsphenoidal surgery for pituitary lesions. Direct closure with a combination of autologous fat, local bone, and/or synthetic grafts remains the standard of care for leaks encountered at the time of surgery as well as postoperatively. The development of the vascularized nasoseptal flap as a closure technique has increased the surgeon’s capacity to correct even larger openings in the dura of the sella as well as widely exposed anterior skull base defects. Yet these advances in the technical nuances for management of post-transsphenoidal CSF leak are useless without the ability to recognize a CSF leak by physical examination, clinical history, biochemical testing, or radiographic assessment. Here, we report a case of a patient who developed a CSF leak 28 years after transsphenoidal surgery, precipitated by a robotic-assisted hysterectomy during which increased intra-abdominal pressure and steep Trendelenberg positioning were both factors. Given the remote nature of the patient’s transsphenoidal surgery and relative paucity of data regarding such a complication, the condition went unrecognized for several months. We review the available literature regarding risk and pathophysiology of CSF leak following abdominal surgery and propose the need for increased vigilance in identification of such occurrences with the increasing acceptance and popularity of minimally invasive abdominal and pelvic surgeries as standards in the field.
IKK-dependent activation of NF-κB contributes to myeloid and lymphoid leukemogenesis by BCR-ABL1
Authors: Hsieh Mo-Ying , Van Etten Richard A.
Published: January 2014

Key Points

Inhibitors of NF-κB activation attenuate lymphoid and myeloid leukemogenesis by BCR-ABL1 and decrease leukemic stem cells in vivo. These results validate IKKs and NF-κB signaling as pharmacological targets for therapy of CML and Ph+ B-ALL.
Gender differences in adiponectin levels and body composition in older adults: Hallym aging study
Authors: Song Hong Ji , Oh Sohee , Quan Shanai , Ryu Ohk-Hyun , Jeong Jin-Young , Hong Kyung-Soon , Kim Dong-Hyun
Published: January 2014

Background

Body composition changes with ageing can influence the adiponectin concentration. However, the component of body composition that is associated with adiponectin concentrations in older adults remains unclear.

Methods

There were 152 males and 168 females aged 65 years or older that participated in the 2010 Hallym Aging Study (HAS). Body composition (assessed by dual energy X-ray absorptiometry; DXA), anthropometric parameters and adiponectin were obtained from all participants. Multivariate linear regression models assessed the association of body fat percentage, regional muscle and bone mineral contents of body composition and waist/height ratio with adiponectin concentration. Age, albumin, testosterone concentration and metabolic parameters were considered as confounding factors.

Results

In correlation analysis, age was positively associated with adiponectin in males (P < 0.01), but not in females. Fasting glucose, albumin, arm skeletal muscle mass and bone mineral content were negatively associated with adiponectin in males (P < 0.05). Testosterone and leg bone mineral content were negatively associated with adiponectin in females (P < 0.05). In multivariate linear regression models, body fat percentage and albumin (P < 0.05) were negatively associated with adiponectin, and high-density lipoprotein cholesterol (HDL-C) (P < 0.001) and age (P < 0.01) were positively associated with adiponectin in older males. In older females, the only factors that correlated significantly with adiponectin concentration were the homeostasis model assessment of insulin resistance (P < 0.001) and HDL-C (P < 0.05). The waist/height ratio and bone mineral content were not associated with adiponectin in either gender.

Conclusion

Plasma adiponectin levels correlated negatively with body fat percentage in older males but not in older females. The differential results between older males and females suggest that certain gender-specific mechanisms may affect the association between adiponectin and age-related body composition changes.
Hypoxia increases membrane metallo-endopeptidase expression in a novel lung cancer ex vivo model – role of tumor stroma cells
Authors: Leithner Katharina , Wohlkoenig Christoph , Stacher Elvira , Lindenmann Jörg , Hofmann Nicole A , Gallé Birgit , Guelly Christian , Quehenberger Franz , Stiegler Philipp , Smolle-Jüttner Freyja-Maria , Philipsen Sjaak , Popper Helmut H , Hrzenjak Andelko , Olschewski Andrea , Olschewski Horst
Published: January 2014

Background

Hypoxia-induced genes are potential targets in cancer therapy. Responses to hypoxia have been extensively studied in vitro, however, they may differ in vivo due to the specific tumor microenvironment. In this study gene expression profiles were obtained from fresh human lung cancer tissue fragments cultured ex vivo under different oxygen concentrations in order to study responses to hypoxia in a model that mimics human lung cancer in vivo.

Methods

Non-small cell lung cancer (NSCLC) fragments from altogether 70 patients were maintained ex vivo in normoxia or hypoxia in short-term culture. Viability, apoptosis rates and tissue hypoxia were assessed. Gene expression profiles were studied using Affymetrix GeneChip 1.0 ST microarrays.

Results

Apoptosis rates were comparable in normoxia and hypoxia despite different oxygenation levels, suggesting adaptation of tumor cells to hypoxia. Gene expression profiles in hypoxic compared to normoxic fragments largely overlapped with published hypoxia-signatures. While most of these genes were up-regulated by hypoxia also in NSCLC cell lines, membrane metallo-endopeptidase (MME, neprilysin, CD10) expression was not increased in hypoxia in NSCLC cell lines, but in carcinoma-associated fibroblasts isolated from non-small cell lung cancers. High MME expression was significantly associated with poor overall survival in 342 NSCLC patients in a meta-analysis of published microarray datasets.

Conclusions

The novel ex vivo model allowed for the first time to analyze hypoxia-regulated gene expression in preserved human lung cancer tissue. Gene expression profiles in human hypoxic lung cancer tissue overlapped with hypoxia-signatures from cancer cell lines, however, the elastase MME was identified as a novel hypoxia-induced gene in lung cancer. Due to the lack of hypoxia effects on MME expression in NSCLC cell lines in contrast to carcinoma-associated fibroblasts, a direct up-regulation of stroma fibroblast MME expression under hypoxia might contribute to enhanced aggressiveness of hypoxic cancers.
Pathology of bladder cancer among diabetic patients undergoing radical cystectomy with a history of pioglitazone (Actos) use
Authors: Romero Victoriano , Peyton Charles , Gray Ian , Hemal Ashok , Terlecki Ryan
Published: January 2014

Background

Prospective studies suggested an association between pioglitazone (Actos) use and the development of bladder cancer. Cancer pathology among pioglitazone users has not been characterized. We chose to compare the surgical pathology among diabetic users and non-users, as well as non-diabetic patients who underwent radical cystectomy for bladder cancer.

Methods

Our single-center, prospectively-maintained bladder cancer database was reviewed. Patient demographics, surgical pathology, and outcomes were evaluated. Information regarding diabetic history and use of pioglitazone was determined from chart analysis and patient interview.

Results

From April 2005 to October 2011, 204 patients undergoing radical cystectomy were identified. Of these, 33 (16.2%) were diabetic and 171 (83.8%) had no history of diabetes. Among diabetic patients, 9 (27.3%) had a history of pioglitazone use. Median duration of therapy was 14 (6–120) months. Pathology in non-diabetic patients was T1 in 17 (9.9%), T2 in 38 (22.2%), T3 in 44 (25.7%), and T4 in 31 (18.1%). Pathology among diabetic non-users was T1 in 1 (4.2%), T2 in 7 (29.2%), T3 in 7 (29.2%), and T4 in 4 (16.7%). Pathologic stage among diabetic users was T1 in 1 (11.1%), T2 in 3 (33.3%), T3 in 3 (33.3%), and T4 in 1 (11.1%). Lymph node involvement in non-diabetics, diabetic non-users, and diabetic users was 25.7%, 33.3%, and 33.3%, respectively. Cancer-specific death was seen in 60.3% of non-diabetics, 58.3% of diabetic non-users, and 75% of diabetic users.

Conclusions

Diabetics have similar stage distribution regardless of pioglitazone use. Lymph node metastases rates and cancer specific death were similar across all groups. Additional studies will serve to better characterize this relationship.
Treatment seeking behaviour in southern Chinese elders with chronic orofacial pain: a qualitative study
Authors: Au Teresa SY , Wong May CM , McMillan Anne S , Bridges Susan , McGrath Colman
Published: January 2014

Background

Chronic orofacial pain (OFP) is common in general adult populations worldwide. High levels of psychological distress and impaired coping abilities are common among Western people with chronic OFP but limited information was found in southern Chinese people. This study aimed to explore the perceptions and experiences of community dwelling elderly people with chronic OFP symptoms and their treatment seeking behaviour in Hong Kong.

Methods

An exploratory qualitative interview study was conducted. Elderly people experiencing chronic OFP symptoms were invited to take part in an individual semi-structured interview. A total of 25 semi-structured interviews were performed for 25 participants.

Results

Pertinent issues relating to the treatment seeking behaviour emerged from the interviews, many of which were inter-related and overlapping. They were organized into three major themes: (i) Impact of chronic OFP on daily life; (ii) Personal knowledge and feeling of chronic OFP; (iii) Management of chronic OFP. The participants were found to have the intention to seek professional treatment, but there were barriers which discouraged them continuing to seek professional treatment. They also received complementary treatment for chronic OFP, such as acupuncture, massage and “chi kung”. Moreover, a wide range self-management techniques were also mentioned. On the other hand, those who did not seek professional treatment for the chronic OFP claimed that they had accepted or adapted to the pain as part of their lives.

Conclusions

This qualitative study observed that elderly people affected by chronic OFP symptoms in Hong Kong sought many different ways to manage their pain including traditional and complementary approaches. The role of the dentist in dealing with chronic OFP is unclear. Multiple barriers exist to accessing care for chronic OFP. The findings may be used to inform future chronic OFP management strategies in Hong Kong.
Assessment of the nursing skill mix in Mozambique using a task analysis methodology
Authors: Dgedge Martinho , Mendoza Angel , Necochea Edgar , Bossemeyer Debora , Rajabo Maharifa , Fullerton Judith
Published: January 2014

Background

The density of the nursing and maternal child health nursing workforce in Mozambique (0.32/1000) is well below the WHO minimum standard of 1 nurse per 1000. Two levels of education were being offered for both nurses and maternal child health nurses, in programmes ranging from 18 to 30 months in length. The health care workforce in Mozambique also includes Medical Technicians and Medical Agents, who are also educated at either basic or mid-level. The Ministry of Health determined the need to document the tasks that each of the six cadres was performing within various health facilities to identify gaps, and duplications, in order to identify strategies for streamlining workforce production, while retaining highest educational and competency standards. The methodology of task analysis (TA) was used to achieve this objective. This article provides information about the TA methodology, and selected outcomes of the very broad study.

Methods

A cross-sectional descriptive task analysis survey was conducted over a 15 month period (2008–2009). A stratified sample of 1295 individuals was recruited from every type of health facility in all of Mozambique’s 10 provinces and in Maputo City. Respondents indicated how frequently they performed any of 233 patient care tasks. Data analysis focused on identifying areas where identical tasks were performed by the various cadres. Analyses addressed frequency of performance, grouped by level of educational preparation, within various types of health facilities.

Results

Task sharing ranged from 74% to 88% between basic and general nurse cadres and from 54% to 88% between maternal and child health nurse cadres, within various health facility types. Conversely, there was distinction between scope of practice for nursing and maternal/child health nursing cadres.

Conclusion

The educational pathways to general nursing and maternal/child health nursing careers were consolidated into one 24 month programme for each career. The scopes of practice were affirmed based on task analysis survey data.
Surface chemistry of SnO2 nanowires on Ag-catalyst-covered Si substrate studied using XPS and TDS methods
Authors: Sitarz Michal , Kwoka Monika , Comini Elisabetta , Zappa Dario , Szuber Jacek
Published: January 2014

In this paper we investigate the surface chemistry, including surface contaminations, of SnO2 nanowires deposited on Ag-covered Si substrate by vapor phase deposition (VPD), thanks to x-ray photoelectron spectroscopy (XPS) in combination with thermal desorption spectroscopy (TDS). Air-exposed SnO2 nanowires are slightly non-stoichiometric, and a huge amount of C contaminations is observed at their surface. After the thermal physical desorption (TPD) process, SnO2 nanowires become almost stoichiometric without any surface C contaminations. This is probably related to the fact that C contaminations, as well as residual gases from air, are weakly bounded to the crystalline SnO2 nanowires and can be easily removed from their surface. The obtained results gave us insight on the interpretation of the aging effect of SnO2 nanowires that is of great importance for their potential application in the development of novel chemical nanosensor devices.
The value of simple microbiological studies for on-site screening of acute neonatal conjunctivitis in Angola
Authors: Alexandre Isabel , Cortes Nestor , Justel Mar , Fernández Itziar , Ortíz de Lejarazu Raul , Pastor J Carlos
Published: January 2014

Background

Neonatal conjunctivitis or ophthalmia neonatorum (ON) is an acute bacterial conjunctivitis contracted by newborns during delivery. In non-industrialized countries, detection of the etiological agent is difficult due to the unavailability of modern diagnostic resources. Therefore, we analyzed the effectiveness of Gram and methylene blue staining techniques, which are simple microbiological methods in suspecting the aetiology of ON in a maternity ward in Luanda, Angola.

Findings

Neonatal conjunctival smears (n = 95), maternal data, and perinatal factors were collected. Slides were air-dried and sent to the Microbiology Department of the Hospital Clinico Universitario, Valladolid, Spain, where trained personnel performed Gram and methylene blue staining methods. Findings were interpreted by two expert microbiologists. Ophthalmological examination of all children showed five newborns with clinical signs of ON. Fourteen mothers reported were suspected with vulvo-vaginitis, and 27 had a urinary infection during pregnancy. Gram staining revealed the presence of epithelial cells in 87.6% and leukocytes in 15% of the conjunctival smears. These values were significantly higher than those shown by methylene blue staining. No rods, cocci, or yeasts were identified by either staining method. Chlamydia trachomatis DNA was also undetected in a small sub-sample with clinical suspicion of ON. There was no correlation among the presence of ON, ON microbes, maternal data, or perinatal factors.

Conclusions

Basic microbiological techniques did not provide enough information for screening cases of ON in Angola. Therefore, the use of molecular biology or other techniques is warranted for this purpose.
Seq and CLIP through the miRNA world
Authors: Mittal Nitish , Zavolan Mihaela
Published: January 2014

High-throughput sequencing of RNAs crosslinked to Argonaute proteins reveals not only a multitude of atypical miRNA binding sites but also of miRNA targets with atypical functions, and can be used to infer quantitative models of miRNA-target interaction strength.
Topoisomerase IIbeta is required for proper retinal development and survival of postmitotic cells
Authors: Li Ying , Hao Hailing , Tzatzalos Evangeline , Lin Ren-Kuo , Doh Sungtae , Liu Leroy F. , Lyu Yi Lisa , Cai Li
Published: January 2014

Topoisomerase IIbeta (Top2b) is an enzyme that modulates DNA supercoiling by catalyzing the passage of DNA duplexes through one another. It is ubiquitously expressed in postmitotic cells and known to function during the development of neuromuscular junctions in the diaphragm and the proper formation of laminar structure in the cerebral cortex. However, due to the perinatal death phenotype of the traditional constitutive and brain-specific Top2b knockout mice, the precise in vivo function of Top2b, especially during postnatal neural development, remains to be determined. Using both the constitutive and retina-specific knockout mouse models, we showed that Top2b deficiency resulted in delayed neuronal differentiation, degeneration of the plexiform layers and outer segment of photoreceptors, as well as dramatic reduction in cell number in the retina. Genome-wide transcriptome analysis by RNA sequencing revealed that genes involved in neuronal survival and neural system development were preferentially affected in Top2b-deficient retinas. Collectively, our findings have indicated an important function of Top2b in proper development and the maintenance/survival of postmitotic neurons in the retina.
Systematic screen for mutants resistant to TORC1 inhibition in fission yeast reveals genes involved in cellular ageing and growth
Authors: Rallis Charalampos , López-Maury Luis , Georgescu Teodora , Pancaldi Vera , Bähler Jürg
Published: January 2014

Target of rapamycin complex 1 (TORC1), which controls growth in response to nutrients, promotes ageing in multiple organisms. The fission yeast Schizosaccharomyces pombe emerges as a valuable genetic model system to study TORC1 function and cellular ageing. Here we exploited the combinatorial action of rapamycin and caffeine, which inhibit fission yeast growth in a TORC1-dependent manner. We screened a deletion library, comprising ∼84% of all non-essential fission yeast genes, for drug-resistant mutants. This screen identified 33 genes encoding functions such as transcription, kinases, mitochondrial respiration, biosynthesis, intra-cellular trafficking, and stress response. Among the corresponding mutants, 5 showed shortened and 21 showed increased maximal chronological lifespans; 15 of the latter mutants showed no further lifespan increase with rapamycin and might thus represent key targets downstream of TORC1. We pursued the long-lived sck2 mutant with additional functional analyses, revealing that the Sck2p kinase functions within the TORC1 network and is required for normal cell growth, global protein translation, and ribosomal S6 protein phosphorylation in a nutrient-dependent manner. Notably, slow cell growth was associated with all long-lived mutants while oxidative-stress resistance was not.
Feminization of pheromone-sensing neurons affects mating decisions in Drosophila males
Authors: Lu Beika , Zelle Kathleen M. , Seltzer Raya , Hefetz Abraham , Ben-Shahar Yehuda
Published: January 2014

The response of individual animals to mating signals depends on the sexual identity of the individual and the genetics of the mating targets, which represent the mating social context (social environment). However, how social signals are sensed and integrated during mating decisions remains a mystery. One of the models for understanding mating behaviors in molecular and cellular terms is the male courtship ritual in the fruit fly (Drosophila melanogaster). We have recently shown that a subset of gustatory receptor neurons (GRNs) that are enriched in the male appendages and express the ion channel ppk23 play a major role in the initiation and maintenance of male courtship via the perception of cuticular contact pheromones, and are likely to represent the main chemosensory pathway that influences mating decisions by males. Here we show that genetic feminization of ppk23-expressing GRNs in male flies resulted in a significant increase in male–male sexual attraction without an apparent impact on sexual attraction to females. Furthermore, we show that this increase in male–male sexual attraction is sensory specific, which can be modulated by variable social contexts. Finally, we show that feminization of ppk23-expressing sensory neurons lead to major transcriptional shifts, which may explain the altered interpretation of the social environment by feminized males. Together, these data indicate that the sexual cellular identity of pheromone sensing GRNs plays a major role in how individual flies interpret their social environment in the context of mating decisions.
Autoregulation of glypican-1 by intronic microRNA-149 fine-tunes the angiogenic response to fibroblast growth factor in human endothelial cells
Authors: Chamorro-Jorganes Aránzazu , Araldi Elisa , Rotllan Noemi , Cirera-Salinas Daniel , Suárez Yajaira
Published: January 2014

MiR-149 is located within the first intron of glypican-1 (GPC1) gene. GPC1 is low affinity receptor for fibroblast growth factor (FGF2) that favors FGF2- binding to its receptor (FGFR1), subsequently promoting FGF2-FGFR1 activation and signaling. Using bioinformatic approaches, both GPC1 and FGFR1 were identified and subsequently validated as targets for miR-149 (both mature and passenger strands) in endothelial cells (ECs). ?As a consequence of their targeting activity towards GPC1 and FGFR1, both miR-149 and miR-149* regulated FGF2 signaling and FGF2-induced responses in ECs, namely proliferation, migration and cord formation. Moreover, lentiviral overexpression of miR-149 reduced in vivo tumor-induced neovascularization. Importantly, FGF2 transcriptionally stimulated the expression of miR-149 independently of its host gene, therefore assuring the steady state of FGF2-induced responses through the regulation of the GPC1-FGFR1 binary complex in ECs.
AII amacrine cells discriminate between heterocellular and homocellular locations when assembling connexin36-containing gap junctions
Authors: Meyer Arndt , Hilgen Gerrit , Dorgau Birthe , Sammler Esther M , Weiler Reto , Monyer Hannah , Dedek Karin , Hormuzdi Sheriar G
Published: January 2014

Electrical synapses (gap junctions) allow for rapid signal transmission between neurons. They are formed by connexins and regulated by a variety of intracellular pathways. In neurons, connexin36 (Cx36) represents the most abundant isoform. The mechanisms forming Cx36-containing electrical synapses are unknown although previous studies indicate that an intact carboxy-terminus is necessary: mice lacking the native Cx36 but expressing a carboxy-terminal fusion variant (KO-Cx36-EGFP) cannot form gap junctions in the olfactory bulb and cerebellum. We now report that neurons may harbor a different mechanism for Cx36 assembly. Using the mouse retina as a model system, we focus on homocellular and heterocellular gap junctions formed by an AII amacrine cell, a key interneuron found in all mammalian retinas. In KO-Cx36-EGFP mice, we demonstrate that heterocellular AII–ON cone bipolar cell gap junctions are fully functional whereas AII–AII homocellular gap junctions are not formed. Tracer injected into an AII amacrine cell spreads into ON cone bipolar cells but is excluded from other AII cells. Reconstructing the set of Cx36-EGFP clusters located on an AII cell in the KO-Cx36-EGFP genotype confirmed that their number but not average size is reduced, as expected for AII cells lacking a subset of electrical synapses. In conclusion, our studies indicate that some neurons exhibit at least two discriminatory mechanisms for assembling Cx36. We suggest that employing different gap junction-forming mechanisms may provide the means for a cell to regulate its gap junctions in a target cell-specific manner even if these junctions contain the same connexin.
Basal localization of MT1-MMP is essential for epithelial cell morphogenesis in 3D collagen matrix
Authors: Weaver Sarah A , Wolters Brit , Ito Noriko , Woskowicz Anna M , Kaneko Kazuyo , Shitomi Yasuyuki , Seiki Motoharu , Itoh Yoshifumi
Published: January 2014

During epithelial tubulogenesis in 3D collagen matrices, the membrane-anchored collagenase MT1-MMP has been shown to play an essential role. However, its regulation during tubulogenesis is not understood. Here we report that degradation of collagen in polarized epithelial cells is post-translationally regulated by changing the localization of MT1-MMP from the apical to the basal surface. Inert polarized epithelial cells predominantly localized MT1-MMP at the apical surface, whereas HGF treatment induced basal localization of MT1-MMP, and resultant collagen degradation. The basal localization of MT1-MMP requires the ecto-domains of the enzyme, as deletion of the MT-Loop region or the hemopexin domain inhibited basal localization of the enzyme. TGFβ is a well-known inhibitor of tubulogenesis and our data indicate that its mechanism of inhibition is at least in part due to inhibition of basal MT1-MMP localization. Interestingly, however, the effect of TGFβ was found to be bi-phasic: it effectively inhibited basal localization at high doses, but promoted basal localization of MT1-MMP and tubulogenesis at lower doses. Taken together these data indicate that basal localization of MT1-MMP is a key factor promoting ECM degradation by polarized epithelial cells, and that this is an essential part of epithelial morphogenesis in 3D collagen.
Epithelial repair is a two-stage process driven first by dying cells and then by their neighbours
Authors: Kuipers Dorothy , Mehonic Aida , Kajita Mihoko , Peter Loïc , Fujita Yasuyuki , Duke Tom , Charras Guillaume , Gale Jonathan E.
Published: January 2014

Epithelial cells maintain an essential barrier despite continuously undergoing mitosis and apoptosis. Biological and biophysical mechanisms have evolved to remove dying cells whilst maintaining that barrier. Cell extrusion is thought to be driven by a multicellular filamentous actin ring formed by the neighbouring cells, with its contraction providing the mechanical force for extrusion, with little or no contribution from the dying cell. We use live confocal imaging, providing time-resolved 3D observations of actomyosin dynamics to reveal new mechanical roles for dying cells in their own extrusion from monolayers. Dying cell clearance could be subdivided into two-stages. The first, previously unidentified, stage was driven by the dying cell, which exerted tension on its neighbours through the action of a cortical contractile F-actin and myosin ring at the cell apex. The second stage, consistent with previous studies, was driven by a multicellular F-actin ring in the neighbouring cells that moved from the apical to the basal plane to extrude the dying cell. Critically, these data reinstate the dying cell as an active physical participant in cell extrusion rather than an innocent bystander.
A central region of Gli2 regulates its localization to the primary cilium and transcriptional activity
Authors: Santos Nicole , Reiter Jeremy F.
Published: January 2014

Signaling through vertebrate Hedgehog (Hh) proteins depends on the primary cilium. In response to Hh signals, the transcriptional activator of the pathway, Gli2, accumulates at the ciliary tip, raising the possibility that ciliary localization is important for Gli2 activation. To test this hypothesis, we used the Floxin system to create knock-in Gli2 alleles in embryonic stem cells (ESCs) that methodically tested which domains and residues are essential for Gli2 ciliary localization. The Gli2 zinc fingers, transcription activation domain, repressor domain, the phosphorylation cluster and a Sufu binding motif were each dispensable for ciliary localization. Mutating residues required for Gli2 sumoylation and nuclear trafficking also did not abrogate ciliary localization. In contrast, several other domains restrict Gli2 nuclear localization, and a central region, distinct from previously characterized domains, was required for ciliary localization. In addition from an inability to localize to cilia, Gli2 lacking this central domain was unable to activate target genes. Thus, our systematic analysis in ESCs reveals that distinct regions of Gli2 regulate its nuclear and ciliary localization. The identification of a domain essential for both ciliary localization and transcriptional activity suggests that ciliary localization of Gli2 is required for its activation.
Quality control of plant peroxisomes in organ specific manner via autophagy
Authors: Yoshimoto Kohki , Shibata Michitaro , Kondo Maki , Oikawa Kazusato , Sato Mayuko , Toyooka Kiminori , Shirasu Ken , Nishimura Mikio , Ohsumi Yoshinori
Published: January 2014

Peroxisomes are essential organelles characterized by the possession of enzymes that produce hydrogen peroxide (H2O2) as part of their normal catalytic cycle. During the metabolic process, peroxisomal proteins are inevitably damaged by H2O2 and the integrity of the peroxisomes is impaired. Here, we show that autophagy, an intracellular process for vacuolar degradation, selectively degrades dysfunctional peroxisomes. Marked accumulation of peroxisomes was observed in the leaves but not roots of autophagy-related (ATG) gene-knockout Arabidopsis thaliana mutants. The peroxisomes in leaf cells contained markedly increased levels of catalase in an insoluble and inactive aggregate form. The chemically inducible complementation system in ATG5 knockout Arabidopsis provided the evidence that these accumulated peroxisomes were delivered to vacuoles by autophagy for degradation. Interestingly, autophagosomal membrane structures specifically recognized the abnormal peroxisomes at the site of the aggregates. Thus, autophagy is essential for the quality control of peroxisomes in leaves for proper plant development under natural growth conditions.
The inverse BAR-domain protein IBARa drives membrane remodelling to control osmoregulation, phagocytosis and cytokinesis
Authors: Linkner Joern , Witte Gregor , Zhao Hongxia , Junemann Alexander , Nordholz Benjamin , Runge-Wollmann Petra , Lappalainen Pekka , Faix Jan
Published: January 2014

Here, we analyzed the single I-BAR family member IBARa from D. discoideum. The X-ray structure of the N-terminal I-BAR domain solved at 2.2 Å resolution revealed an all-α helical structure that self-associates into a 165 Å zeppelin-shaped antiparallel dimer. The structural data are consistent with its shape in solution obtained by small-angle X-ray-scattering. Cosedimentation, fluorescence-anisotropy as well as fluorescence and electron microscopy revealed the I-BAR domain to bind preferentially to phosphoinositide-containing vesicles and drive the formation of negatively curved tubules. Immunofluorescence labelling further showed accumulation of endogenous IBARa at the tips of filopodia, the rim of constricting phagocytic cups, in foci connecting dividing cells during the final stage of cytokinesis, and most prominently at the osmoregulatory contractile vacuole (CV). Consistently, IBARa-null mutants displayed defects in CV formation and discharge, growth, phagocytosis and mitotic cell division, whereas filopodia formation was not compromised. Of note, IBARa-null mutants were also strongly impaired in cell spreading. Together, these data suggest IBARa to constitute an important regulator of numerous cellular processes intimately linked with the dynamic rearrangement of cellular membranes.
An ancestral non-proteolytic role for presenilin proteins in multicellular development of the social amoeba Dictyostelium discoideum
Authors: Ludtmann Marthe H. R. , Otto Grant P. , Schilde Christina , Chen Zhi-Hui , Allan Claire Y. , Brace Selina , Beesley Philip W. , Kimmel Alan R. , Fisher Paul , Killick Richard , Williams Robin S. B.
Published: January 2014

Mutations in two presenilin genes cause familial Alzheimer's disease. Presenilins have both proteolysis-dependent functions as components of the γ-secretase complex and proteolysis-independent functions in signalling. In this study we investigate a conserved function of human presenilins in the development of the simple model Dictyostelium discoideum. We show that the block in Dictyostelium development caused by ablation of both Dictyostelium presenilins is rescued by expression of human presenilin 1, restoring terminal differentiation of multiple cell types. This developmental role is independent of proteolytic activity, since mutation of both catalytic aspartates does not affect presenilin ability to rescue development, and ablation of nicastrin, a γ-secretase component critical for proteolytic activity, does not block development. The role of presenilins during Dictyostelium development is therefore independent of their proteolytic activity. However, presenilin loss in Dictyostelium results in elevated cyclic AMP (cAMP) levels and enhanced stimulation-induced calcium release, suggesting that presenilins regulate these intracellular signalling pathways. Our data suggest that presenilin proteins perform an ancient non-proteolytic role in regulating intracellular signalling and development, and that Dictyostelium is a useful model for analysing human presenilin function.
Modulation of hypoxia-signaling pathways by extracellular long non-coding RNA regulator of reprogramming
Authors: Takahashi Kenji , Yan Irene K. , Haga Hiroaki , Patel Tushar
Published: January 2014

Resistance to adverse environmental conditions such as hypoxia contributes to the efficacy of anticancer therapies and tumor progression. Although deregulated expression of many long non-coding RNA (lncRNA) occur in human cancers, their contributions to tumor responses to hypoxia are unknown. RNA expression profiling identified several hypoxia-responsive lncRNAs including linc-RoR, which is also increased in expression in malignant liver cancer cells. Linc-RoR expression was increased in hypoxic regions within tumor cell xenografts in vivo. Tumor cell viability during hypoxia was reduced by siRNA to linc-RoR. Compared to controls, siRNA to linc-RoR decreased p70S6K1 phosphorylation, PDK1 and HIF-1α protein expression, and increased expression of the linc-RoR target miR-145. Linc-RoR was highly expressed in extracellular RNA released by HCC cells during hypoxia. Incubation with extracellular vesicle preparations containing extracellular RNA increased linc-RoR, HIF-1α expression and cell survival in recipient cells. These studies show that lncRNA-RoR is a hypoxia-responsive lncRNA that is functionally linked to hypoxia signaling in HCC through a miR-145/HIF-1α signaling module, and identify a mechanistic role of inter-cellular extracellular transfer of linc-RoR in promoting cell survival during hypoxic stress.
Mechanistic comparison of Bacillus subtilis 6S-1 and 6S-2 RNAs—commonalities and differences
Authors: Burenina Olga Y. , Hoch Philipp G. , Damm Katrin , Salas Margarita , Zatsepin Timofei S. , Lechner Marcus , Oretskaya Tatiana S. , Kubareva Elena A. , Hartmann Roland K.
Published: January 2014

Bacterial 6S RNAs bind to the housekeeping RNA polymerase (σA-RNAP in Bacillus subtilis) to regulate transcription in a growth phase-dependent manner. B. subtilis expresses two 6S RNAs, 6S-1 and 6S-2 RNA, with different expression profiles. We show in vitro that 6S-2 RNA shares hallmark features with 6S-1 RNA: Both (1) are able to serve as templates for pRNA transcription; (2) bind with comparable affinity to σA-RNAP; (3) are able to specifically inhibit transcription from DNA promoters, and (4) can form stable 6S RNA:pRNA hybrid structures that (5) abolish binding to σA-RNAP. However, pRNAs of equal length dissociate faster from 6S-2 than 6S-1 RNA, owing to the higher A,U-content of 6S-2 pRNAs. This could have two mechanistic implications: (1) Short 6S-2 pRNAs (<10 nt) dissociate faster instead of being elongated to longer pRNAs, which could make it more difficult for 6S-2 RNA-stalled RNAP molecules to escape from the sequestration; and (2) relative to 6S-1 RNA, 6S-2 pRNAs of equal length will dissociate more rapidly from 6S-2 RNA after RNAP release, which could affect pRNA turnover or the kinetics of 6S-2 RNA binding to a new RNAP molecule. As 6S-2 pRNAs have not yet been detected in vivo, we considered that cellular RNAP release from 6S-2 RNA might occur via 6S-1 RNA displacing 6S-2 RNA from the enzyme, either in the absence of pRNA transcription or upon synthesis of very short 6S-2 pRNAs (∼5-mers, which would escape detection by deep sequencing). However, binding competition experiments argued against these possibilities.
Screening Method to Evaluate Point-of-Care Human Chorionic Gonadotropin (hCG) Devices for Susceptibility to the Hook Effect by hCG β Core Fragment: Evaluation of 11 Devices
Authors: Nerenz Robert D , Song Haowei , Gronowski Ann M
Published: January 2014

BACKGROUND

The predominant hCG variant in urine, hCG β core fragment (hCGβcf), has been demonstrated to cause false-negative results in qualitative point-of-care (POC) hCG devices. This is a major concern for healthcare professionals using POC pregnancy tests. We developed a screening method to evaluate qualitative POC hCG devices for their susceptibility to inhibition by hCGβcf. Using this method, we evaluated the performance of 11 commonly used devices.

METHODS

A wide range of purified hCG and hCGβcf concentrations were mixed and tested on 2 POC devices. By use of those results, a screening method was defined and 9 additional POC devices were evaluated. Two solutions containing (a) 500 pmol/L (171 IU/L) intact hCG with 0 pmol/L hCGβcf and (b) 500 pmol/L intact hCG with 500 000 pmol/L hCGβcf were used to screen all POC devices.

RESULTS

The OSOM and Cen-Med Elite devices were found to be most susceptible to false-negative results due to hCGβcf. The BC Icon 20 and the Alere were the least susceptible. The remaining 7 were moderately affected. Devices that gave the strongest signal with hCGβcf alone were those that were least likely to show a hook effect.

CONCLUSIONS

The screening method put forth here can be used by device users and manufacturers to evaluate POC devices for inhibition by hCGβcf. Of 11 devices evaluated, only 2 have been identified that exhibit minimal to no susceptibility to hCGβcf.
The contribution of fat component to gestational weight gain
Authors: Pokusaeva V N , Troshina E A , Nikiforovskaya E N , Vahrushina A S
Published: January 2014

Objective: to estimate the role of adipose tissue in gestational weight gain (GWG) and preferential fat deposition among normal-weight women. Subjects and methods: prospective cohort study of 84 pregnancies: maternal body mass index 18,5–24,9 kg/m2, singleton term pregnancy, nondiabetic women, somatically well. GWG and skinfold thickness were evaluated in the 1st, 2nd, 3d trimesters, on the 3d day after delivery. Results: fat mass gain in low GWG was similar to recommended GWG and in the high-GWG group was greater one. Women with recommended and low GWG returned to their initial fat level on the 3d day after delivery, in excessive weight gain fat significantly increased (р=0,025). Compared to initial recommended GWG resulted in triceps skinfold thicknesses loss (р=0,001), in abdominal skinfold gained nothing and in thighs skinfold thicknesses increasing (р=0,021). Inadequate GWG leads to fat loss in arms (р=0,017), fat of abdominal area and thighs return to initial level. In excessive GWG fat in the upper trunk and arms not changed, in the lower area (thighs) significantly increased compared to initial level (р=0,001) or other groups (р=0,001). Conclusion: excessive GWG was associated with greater adipose tissue cumulation and its deposition preferentially over the thighs. Inadequate GWG was clearly linked to low fat-free mass gain.
Features of glucose metabolism in acromegaly depending on the type of treatment
Authors: Dreval A V , Trigolosova I V , Vinogradova A V , Ilovaiskaya I A , Tisheninova R S
Published: January 2014

Aim of the study was to investigate the features of glucose metabolism in patients with acromegaly depending on treatment. Materials and methods: 63 patients with acromegaly: 29 patients with newly diagnosed acromegaly, 25 patients receiving somatostatin analogs (SSA) therapy and 9 patients underwent surgical treatment. Patients with earlier revealed diabetes mellitus (DM) were not included. The characterization of glucose metabolism was carried out by means of an assessment of fasting insulin plasma levels (FIP) and indexes of an insulinresistance (IR) HOMA-R and an insulinsensivity (IS) MATSUDA. Results: In patients receiving SSA therapy the carbohydrate metabolism disorders (CMD) were revealed in 92%, among newly diagnosed acromegalic patients – in 62% and in patients after surgical treatment in 33% of cases. In newly diagnosed acromegalic patients index MATSUDA was 3,5 times lower than in patients receiving SSA therapy, and 4 times lower than in patients after surgical treatment ((р=0,001). НОМА-R in newly diagnosed acromegalic patients was 3,7 times higher than in other groups, p=0,003 and the level of FIP was 4,3 times higher than in patients receiving SSA therapy and 2,9 times higher than in patients after surgical treatment (р=0,001). The index MATSUDA in newly diagnosed acromegalic patients with early CMD was 2,9 times lower, р=0,006, НОМА-R – 2,5 times higher, р=0,025 and FIP 2,4 times higher, р=0,039 than in patients with early CMDs without acromegaly. Conclusions: In patients with newly diagnosed acromegaly in the absence of CMDs increase in IR is compensated by hyperinsulinemia. In patients, receiving SSA therapy the IR decreases, however suppression of secretion of insulin leads to increase in percentage of DM in this group. After surgical treatment IR decreases against higher level of fasting plasma insulin that leads to normalization of carbohydrate metabolism. Keywords: Acromegaly, secondary diabetes mellitus, insulinresistance, MATSUDA, HOMA-R.
On the surgical treatment of the metabolic syndrome
Authors: Sedletsky Y I , Neymark A E , Mirchuk K K , Anisimova K A
Published: January 2014

We present he results of surgical treatment of 136 patients with metabolic syndrome at long-term period of evaluation (up to 15 years). All patients had jejunoileal bypass surgery in modification prof. Y.I. Sedletsky. We show the effect of surgery on overweight, hypertension, dyslipidemia, and hyperglycemia. We've traced above changes depending on the period elapsed since the time of the operation. The results prove the efficiency and stability of effects of jejunoileal bypass surgery on components of the metabolic syndrome. Evaluation of the complications rate of this method is also presented in the article.
Obesity Paradox" – another look at the problem of cardiovascular disease
Authors: Shpagina O V , Bondarenko I Z
Published: January 2014

Major epidemiologic studies over the last century demonstrated that obesity leads to several severe diseases such as diabetes mellitus, hypertension, coronary heart disease, chronic heart failure, cerebrovascular accidents. In developed countries cardiovascular diseases became the main cause of death. In the last 5–6 years some studies showed that people with overweight and obesity of the first degree have a higher life expectancy than people with normal weight. In 2009, the published data showed that the presence of obesity in patients with chronic heart failure does not impair cardiovascular prognosis. Overweight correlates with a decrease in overall mortality by 25%. And in a first degree of obesity the risk of death is reduced by 12%. This phenomenon is called "obesity paradox" and the causes of which are discussed in this review.