Lipophilic Statins Inhibit YAP Nuclear Localization, Coactivator Activity, and Migration in Response to Ligation of HLA Class I Molecules in Endothelial Cells: Role of YAP Multisite Phosphorylation.

Solid-organ transplant recipients exhibiting HLA donor-specific Abs are at risk for graft loss due to chronic Ab-mediated rejection. HLA Abs bind HLA molecules expressed on the surface of endothelial cells (ECs) and induce intracellular signaling pathways, including the activation of the transcriptional coactivator yes-associated protein (YAP). In this study, we examined the impact of lipid-lowering drugs of the statin family on YAP localization, multisite phosphorylation, and transcriptional activity in human ECs. Exposure of sparse cultures of ECs to cerivastatin or simvastatin induced striking relocalization of YAP from the nucleus to the cytoplasm and inhibited the expression of the YAP/TEA domain DNA-binding transcription factor-regulated genes connective tissue growth factor and cysteine-rich angiogenic inducer 61. In dense cultures of ECs, statins prevented YAP nuclear import and expression of connective tissue growth factor and cysteine-rich angiogenic inducer 61 stimulated by the mAb W6/32 that binds HLA class I. Exposure of ECs to either cerivastatin or simvastatin completely blocked the migration of ECs stimulated by ligation of HLA class I. Exogenously supplied mevalonic acid or geranylgeraniol reversed the inhibitory effects of statins on YAP localization either in low-density ECs or high-density ECs challenged with W6/32. Mechanistically, cerivastatin increased the phosphorylation of YAP at Ser127, blunted the assembly of actin stress fiber, and inhibited YAP phosphorylation at Tyr357 in ECs. Using mutant YAP, we substantiated that YAP phosphorylation at Tyr357 is critical for YAP activation. Collectively, our results indicate that statins restrain YAP activity in EC models, thus providing a plausible mechanism underlying their beneficial effects in solid-organ transplant recipients.

Ligation of HLA Class I Molecules Induces YAP Activation through Src in Human Endothelial Cells.

Ab cross-linking of HLA class I (HLA I) molecules on the surface of endothelial cells (EC) triggers proliferative and prosurvival intracellular signaling, which is implicated in the process of chronic allograft rejection, also known as transplant vasculopathy. Despite the importance of Ab-mediated rejection in transplantation, the mechanisms involved remain incompletely understood. In this study, we examined the regulation of yes-associated protein (YAP) localization, phosphorylation, and transcriptional activity in human ECs challenged with Abs that bind HLA I. In unstimulated ECs, YAP localized mainly in the cytoplasm. Stimulation of these cells with Ab W6/32 induced marked translocation of YAP to the nucleus. The nuclear import of YAP was associated with a rapid decrease in YAP phosphorylation at Ser127 and Ser397, sites targeted by LATS1/2 and with the expression of YAP-regulated genes, including connective tissue growth factor (CTGF), and cysteine-rich angiogenic inducer 61 (CYR61). Transfection of small interfering RNAs targeting YAP/TAZ blocked the migration of ECs stimulated by ligation of HLA I, indicating that YAP mediates the increase in EC migration induced by HLA I ligation. Treatment of intact ECs with Src family inhibitors induced cytoplasmic localization of YAP in unstimulated ECs and, strikingly, blocked the nuclear import of YAP induced by Ab-induced HLA I activation in these cells and the increase in the expression of the YAP-regulated genes CTGF and CYR61 induced by HLA I stimulation. Our results identify the Src/YAP axis as a key player in promoting the proliferation and migration of ECs that are critical in the pathogenesis of transplant vasculopathy.

Differentially regulated gene expression in quiescence versus senescence and identification of ARID5A as a quiescence associated marker.

In multicellular organisms majority of the cells remain in a non-dividing states of either quiescence (reversible) or senescence (irreversible). In the present study, gene expression signatures unique to quiescence and senescence were identified using microarray in osteosarcoma cell line, U2OS. It was noted that certain genes and pathways like NOD pathway was shared by both the growth arrest conditions. A major highlight of the present study was increased expression of number of chemokines and cytokines in both quiescence and senescence. While senescence-associated secretory phenotype (SASP) is well known, the quiescence-associated secretory phenotype (QASP) is relatively unknown and appeared novel in this study. ARID5A, a subunit of SWI/SNF complex was identified as a quiescence associated gene. The endogenous expression of ARID5A increased during serum starved condition of quiescence. Overexpression of ARID5A resulted in more number of cells in G0/G1 phase of cell cycle. Further ARID5A overexpressing cells when subjected to serum starvation showed a pronounced secretory phenotype. Overall, the present work has identified gene expression signatures which can distinguish quiescence from senescence.

Growth arrest of lung carcinoma cells (A549) by polyacrylate-anchored peroxovanadate by activating Rac1-NADPH oxidase signalling axis.

Hydrogen peroxide is often required in sublethal, millimolar concentrations to show its oxidant effects on cells in culture as it is easily destroyed by cellular catalase. Previously, we had shown that diperoxovanadate, a physiologically stable peroxovanadium compound, can substitute H2O2 effectively in peroxidation reactions. We report here that peroxovanadate when anchored to polyacrylic acid (PAPV) becomes a highly potent inhibitor of growth of lung carcinoma cells (A549). The early events associated with PAPV treatment included cytoskeletal modifications, increase in GTPase activity of Rac1, accumulation of the reactive oxygen species, and also increase in phosphorylation of H2AX (γH2AX), a marker of DNA damage. These effects persisted even at 24 h after removal of the compound and culminated in increased levels of p53 and p21 together with growth arrest. The PAPV-mediated growth arrest was significantly abrogated in cells pre-treated with the N-acetylcysteine, Rac1 knocked down by siRNA and DPI an inhibitor of NADPH oxidase. In conclusion, our results show that polyacrylate derivative of peroxovanadate efficiently arrests growth of A549 cancerous cells by activating the axis of Rac1-NADPH oxidase leading to oxidative stress and DNA damage.

Copper-phenanthroline catalysts for regioselective synthesis of pyrrolo[3',4':3,4]pyrrolo[1,2-a]furoquinolines/phenanthrolines and of pyrrolo[1,2-a]phenanthrolines under mild conditions.

A new series of pyrrolo[3',4':3,4]pyrrolo[1,2-a]furoquinolines/phenanthrolines and pyrrolo[1,2-a]phenanthrolines were efficiently built up from an 8-hydroxyquinoline derivative or phenanthroline via 1,3-dipolar cycloaddition reaction involving non-stabilized azomethine ylides, generated in situ from the parent furo[3,2-h]quinoliniums/phenanthroliums, in presence of a copper(II) chloride-phenanthroline catalytic system. The methodology combines general applicability with high yields.

Light-Intensity Switching of Graphene/WSe2 Synaptic Devices.

2D van der Waals heterojunctions (vdWH) have emerged as an attractive platform for the realization of optoelectronic synaptic devices, which are critical for energy-efficient computing systems. Photogating induced by charge traps at the interfaces indeed results in ultrahigh responsivity and tunable photoconductance. Yet, optical potentiation and depression remain mostly modulated by gate bias, requiring relatively high energy inputs. Thus, advanced all-optical synapse switching strategies are still needed. In this work, a reversible switching between positive photoconductivity (PPC) and negative photoconductivity (NPC) is achieved in graphene/WSe2 vdWH solely through light-intensity modulation. Consequently, the graphene/WSe2 synaptic device shows tunable optical potentiation and depression behavior with an ultralow power consumption of 127 aJ. The study further unravels the complex interplay of gate bias and incident light power in determining the sign and magnitude of the photocurrent, showing the critical role of charge trapping and photogating at interfaces. Interestingly, it is found that switching between PPC to NPC can be also obtained at 0 mV drain-source voltage. Overall, the reversible potentiation/depression effect based on light intensity modulation and its combination with additional gate bias tunability is very appealing for the development of energy-efficient optical communications and neuromorphic computing.

Opposite Effects of Src Family Kinases on YAP and ERK Activation in Pancreatic Cancer Cells: Implications for Targeted Therapy.

Pancreatic ductal adenocarcinoma (PDAC) remains an aggressive disease that is expected to become the second cause of cancer fatalities during the next decade. As therapeutic options are limited, novel targets, and agents for therapeutic intervention are urgently needed. Previously, we identified potent positive crosstalk between insulin/IGF-1 receptors and G protein-coupled (GPCR) signaling systems leading to mitogenic signaling in PDAC cells. Here, we show that a combination of insulin and the GPCR agonist neurotensin induced rapid activation of Src family of tyrosine kinases (SFK) within PANC-1 cells, as shown by FAK phosphorylation at Tyr576/577 and Tyr861, sensitive biomarkers of SFK activity within intact cells and Src416 autophosphorylation. Crucially, SFKs promoted YAP nuclear localization and phosphorylation at Tyr357, as shown by using the SFK inhibitors dasatinib, saracatinib, the preferential YES1 inhibitor CH6953755, siRNA-mediated knockdown of YES1, and transfection of epitogue-tagged YAP mutants in PANC-1 and Mia PaCa-2 cancer cells, models of the aggressive squamous subtype of PDAC. Surprisingly, our results also demonstrate that exposure to SFK inhibitors, including dasatinib or knockdown of YES and Src induces ERK overactivation in PDAC cells. Dasatinib-induced ERK activation was completely abolished by exposure to the FDA-approved MEK inhibitor trametinib. A combination of dasatinib and trametinib potently and synergistically inhibited colony formation by PDAC cells and suppressed the growth of Mia PaCa-2 cells xenografted into the flank of nude mice. The results provide rationale for considering a combination(s) of FDA-approved SFK (dasatinib) and MEK (e.g., trametinib) inhibitors in prospective clinical trials for the treatment of PDAC.

Critical measurement issues in the assessment of social media influence on body image.

Progress towards understanding how social media impacts body image hinges on the use of appropriate measurement tools and methodologies. This review provides an overview of common (qualitative, self-report survey, lab-based experiments) and emerging (momentary assessment, computational) methodological approaches to the exploration of the impact of social media on body image. The potential of these methodologies is detailed, with examples illustrating current use as well as opportunities for expansion. A key theme from our review is that each methodology has provided insights for the body image research field, yet is insufficient in isolation to fully capture the nuance and complexity of social media experiences. Thus, in consideration of gaps in methodology, we emphasise the need for big picture thinking that leverages and combines the strengths of each of these methodologies to yield a more comprehensive, nuanced, and robust picture of the positive and negative impacts of social media.

Hydroelectric power plant on a paper strip.

We exploit the combinatorial advantage of electrokinetics and tortuosity of a cellulose-based paper network on laboratory grade filter paper for the development of a simple, inexpensive, yet extremely robust (shows constant performance for 12 days) 'paper-and-pencil'-based device for energy harvesting applications. We successfully achieve harvesting of a maximum output power of ∼640 pW in a single channel, while the same is significantly improved (by ∼100 times) with the use of a multichannel microfluidic array (maximum of up to 20 channels). Furthermore, we also provide theoretical insights into the observed phenomenon and show that the experimentally predicted trends agree well with our theoretical calculations. Thus, we envisage that such ultra-low cost devices may turn out to be extremely useful in energizing analytical microdevices in resource limited settings, for instance, in extreme point of care diagnostic applications.

Increased expression of SIRT2 is a novel marker of cellular senescence and is dependent on wild type p53 status.

Sirtuins (SIRT) belonging to the NAD+ dependent histone deacetylase III class of enzymes have emerged as master regulators of metabolism and longevity. However, their role in prevention of organismal aging and cellular senescence still remains controversial. In the present study, we now report upregulation of SIRT2 as a specific feature associated with stress induced premature senescence but not with either quiescence or cell death. Additionally, increase in SIRT2 expression was noted in different types of senescent conditions such as replicative and oncogene induced senescence using multiple cell lines. Induction of SIRT2 expression during senescence was dependent on p53 status as depletion of p53 by shRNA prevented its accumulation. Chromatin immunoprecipitation revealed the presence of p53 binding sites on the SIRT2 promoter suggesting its regulation by p53, which was also corroborated by the SEAP reporter assay. Overexpression or knockdown of SIRT2 had no effect on stress induced premature senescence, thereby indicating that SIRT2 increase is not a cause of senescence; rather it is an effect linked to senescence-associated changes. Overall, our results suggest SIRT2 as a promising marker of cellular senescence at least in cells with wild type p53 status.

Sirtuin 7 in cell proliferation, stress and disease: Rise of the Seventh Sirtuin!

Sirtuin 7 is a member of the sirtuin family of proteins. Sirtuins were originally discovered in yeast for its role in prolonging replicative lifespan. Until recently SIRT7 happened to be the least studied sirtuin of the seven mammalian sirtuins. However, a number of recent breakthrough reports have provided significant clarity to SIRT7 biology. SIRT7 is now seen as a vital regulator of rRNA and protein synthesis for maintenance of normal cellular homeostasis. Proteins like p53, H3K18, PAF53, NPM1 and GABP-ß1 are the known substrates for the deacetylase activity of SIRT7, thereby making it a key mediator of many cellular activities. Studies using in vitro based assays and also knockout mice have revealed a role of SIRT7 in certain disease pathologies as well. High expression of SIRT7 has been reported in few cancer types and is steadily propelling SIRT7 towards an oncogene status. The role of SIRT7 as a pro-survival adaptor molecule in conditions of cellular stress has recently emerged in view of the fact that SIRT7 can regulate molecules like HIF and IRE1α. Additionally, SIRT7 plays a key role in maintenance of the epigenome as it caused the deacetylation of histone (H3K18) and global proteomics studies have shown its interaction with many chromatin remodelling complexes such as B-WICH and other proteins. Lately, the role of SIRT7 in hepatic lipid metabolism has been debated. This review attempts to summarize these recent findings and present the role of SIRT7 as an important cellular regulator.

Role of cellular senescence in hepatic wound healing and carcinogenesis.

A state of permanent growth arrest characterises a senescent cell. Both the beneficial and deleterious effects that have accrued in senescent cells are observed in a complex organ, such as the liver. Injury to liver tissues triggers processes of regeneration and associated wound healing. Persistent injury can also lead to the neoplastic state. Recent evidence linked the senescent characteristics of the cells to the beneficial processes of wound healing and tumour surveillance in the liver. On the other hand, the secretory phenotype of senescent cells can also selectively promote undesirable neoplastic progression. In an evolutionary context, a senescent cell can function primarily as an adaptive response featuring the characteristics of altruism, trade-offs and bystander effects. Using the liver cell as a model system, this review focuses on the current knowledge of the role of senescence in these seemingly contradictory cell phenomena.

A study of surgical management of chronic suppurative otitis media with cholesteatoma and its outcome.

OBJECTIVE: Aim of this study is evaluation of course of improvement of surgically treated cases of chronic suppurative otitis media (CSOM) with cholesteatoma; it includes hearing status, condition of mastoid cavity, study of different, natural and surgical condition and recurrence of disease within the study period. DESIGN: It is a prospective study. SETTINGS: This study was conducted in a premiere government hospital in Kolkata between May 2007 to April 2008. PATIENTS: Total 40 patients between age group of 6-70 years were included in the present study which includes 19 males and 21 females. INTERVENTION: Surgical interventions were done in all the cases. Different types of mastoidectomy with or without tympanoplasty was done according to extent of disease process. OUTCOME: Audiometrically documentable hearing improvement occurred in 35% cases (p = 14), in rest of the ears hearing status remained unaltered. At the end of 6 months follow up 92.5% (p = 14) in rest (p = 37) operated ears become completely dry. Five percent cases (p = 2) presented with facial paralysis; among them one patient improved completely and another patient improved from grade V to grade III facial paralysis. No patient developed any post operative intracranial complications and recurrence of cholesteatema not found in 6 months follow up. Meatal stenosis developed in 5% cases (p = 2) at the end of 6 months. CONCLUSION: Surgery is mainstay of treatment in CSOM with cholesteatoma. Eradication of disease, prevention of complication, maintenance and restoration of hearing, and giving the patient a non-discharging ear are main aim of treatment.

An arrow penetrating at base of the skull successfully removed.

A 30-year-old male presented with accidental injury with an arrow which referred to us from a peripheral village hospital. It was found that the arrow was penetrating through the nasal bones. An xray skull lateral view showed the tip of the arrow penetrating into the posterior wall of the sphenoid sinus. As the patient had no clinical evidence of neurological or vascular injury, he was immediately operated upon and the arrow was removed. Patient was discharged in good condition and a 3-month follow-up was normal.

Granular cell myoblastoma of the tongue in a 2-year-old girl: a case report.

Granular cell tumours are uncommon lesions, although the head and neck region accounts for approximately 50% of all lesions. It is not clear whether or not granular cell tumour is a true neoplasm, a developmental anomaly, or a trauma-induced proliferation. The basic cell of origin is now thought to be neural, although past reports frequently indicated an origin from striated muscle, or less frequently an origin from histiocytes, fibroblasts or pericytes. The tongue and the buccal mucosa are common intraoral sites. The other head and neck site likely to be involved is the larynx. The tumour generally occurs in middle or older aged adults. More than a third of all granular cell tumours occur on the lingual dorsum, usually as a sessile, painless, somewhat firm, immoveable nodule less than 1.5 cm in greatest diameter. Lesions often demonstrate a pallor or a yellowish discolouration and typically have a smooth surface. Histochemical and ultrastructural studies propose the origin of the lesion from Schwann cells, striated muscle, mesenchymal cells, histiocytes and epithelial cells. As most of the granular cell tumours are benign, surgical excision of the lesion is the treatment of choice.

Clinicopathological study of otogenic brain abscess.

Chronic suppurative otitis media (CSOM) is one of the commonest disease entities encountered in otolaryngology practice. Due to poor economic conditions poor hygiene lack of education and death of knowledge about the disease and its complications, we frequently encounter patients of CSOM with extracranial and intracranial complications. Among the intracranial complications otogenic brain abscess is one of the dreadest. With the improvement of healthcare and accessibility of the poor people to healthcare system and development of CT scan, MRI as diagnostic tools incidence of otogenic brain abscess is becoming less due to early diagnosis and interventions. The objectives of this study are to assess the incidence of brain abscesses to explore age, sex, variation to study clinical presentations and common pathogens responsible.