Characterization of Fut10 and Fut11, Putative Alpha-1-3/4 Fucosyltransferase Genes Important for Vertebrate Development

Two new, putative alpha-1-3/4 fucosyltransferases ([alpha]1-3/4 Fuc-Ts), Fuc-TX and Fuc-TXI, were identified in the vertebrate genome and transcriptome sequence databases through sequence homology-based queries. These proteins have a significant sequence similarity to only [alpha]1-3/4 Fuc-Ts, and possess peptide motifs that are evolutionarily conserved among the known vertebrate [alpha]1-3/4 Fuc-Ts. However, Fuc-TX and Fuc-TXI lack the HH[R/W][D/E] sequence that determines the specificity for type 1 or 2 substrates among the known vertebrate enzymes, and Fuc-TXI proteins do not possess a transmembrane domain. The Fut10 and Fut11 genes that encode these proteins are expressed ubiquitously in the adult mouse and in the mouse embryo throughout development. Though a Fuc-T activity of the mouse proteins could not be detected, Fuc-TXI, but not Fuc-TX, was found to hydrolyze GDP-fucose. The interaction of Fuc-TXI with GDP-fucose was also confirmed by its binding to GDP-hexanolamine. In zebrafish, Fut11 transcripts could be detected during early embryonic development. A knock-down of Fuc-TXI in zebrafish embryos with Fut11-specific antisense morpholino oligonucleotides resulted in malformations of the posterior trunk and tail

Detection of MicroRNAs in Dried Serum Blots

MicroRNAs are short RNAs which can be utilized as biomarkers for a variety of conditions. They are detectable in serum, and changes in the levels of circulating microRNAs have been associated with different diseases. We tested for the presence of microRNAs in serum that is dried on paper and stored unrefrigerated instead of being kept frozen. MicroRNAs were readily detectable in such blots, and their detectability was improved by using paper made of cellulose instead of glass fiber, and by re-hydrating dried blots with Trizol™ instead of water, phosphate-buffered saline, or guanidine hydrochloride before RNA extraction. MicroRNA preservation was not diminished by drying of blots at 37, 45 or 60 ºC instead of room temperature or by storage of blots at 37, 45 or 60 ºC instead of room temperature, but was worse when blots were dried incompletely or exposed to high humidity during storage. Preservation of microRNAs in serum was not adversely affected if instead of being kept frozen at -80 ºC it was stored as dried blots at room temperature or 37 ºC for ten or eight and a half months, respectively. In a group of ten individuals, microRNA quantifications of -80 ºC-frozen or dried sera stored at room temperature correlated well. Dried blots may thus be a convenient and safer way to save, transport, and store serum without refrigeration for microRNA assays

Intersubjectivity World of Virtual Reality: Facebook Users Behaviour in the Context of Privacy, Self and Identity

This paper brings together the perspective of social production of space as a 'category' which creates the 'inter-subjective' world of virtual reality where interaction is subjected to individual's virtual 'privacy' and in turn shapes their extended 'self and identity'. This reality keeps on negotiating with the actor as an external entity which can be identified as 'identity' and also as an internal entity which is often categorised as 'self'. The perspectives on extension of space blurred the notion of reality and imagination by identifying itself with Baudrillard's (1983) notion of 'hyper-reality'. And this is being considered to study the 'intersubjectivity' of virtual reality. Empirically, the paper focuses on young Facebook user's behaviour to understand major questions like young users concern on the issue of 'privacy' in virtual space and their presentation of 'self and identity'. For the construction of the young user's notion on "privacy, self and identity", we have considered the qualitative explanation provided by 80 young users across different parts of the countr

Overexpression of MicroRNA miR-30a or miR-191 in A549 Lung Cancer or BEAS-2B Normal Lung Cell Lines Does Not Alter Phenotype

alterations in human lung cancer. compared to their controls. does not lead to an alteration in cell cycle, proliferation, xenograft formation, and chemosensitivity of A549 and BEAS-2B cell lines. Using microarray data from whole tumors to select specific miRNAs for functional study may be a suboptimal strategy

What Does Increasing Labour Homogeneity Mean for Indian Manufacturing?

In evaluating the economic system, the share of wage in value-added (labour share) assumes significance. We examine the trend of labour share in Indian manufacturing during 2001-14, by using the pooled plant-level data. In the analysis, the heterogeneities with respect to factor share, factor ratio and magnitude of factors (labour and capital) are gauged, while the trend of elasticity of substitution is measured. With this context, we assess if the labour heterogeneity explains the variation in the total factor productivity, taking the industry as a unit of analysis. Further, the analysis looks into the relationship between labour heterogeneity and productivity. Analysing the microdata on the labour force, we gauge the determinants of wage from the vantage of supply. Cues from this analysis point to the need for social up-gradation of Indian manufacturing in terms of decent employment relations and skill. This change may enable India to move from factor abundant system to a productivity-oriented economy in the milieu of steadfast substitution of labour by capital. The novelty of this paper is in pooling the plant-level data across years, while industry level aggregation is resorted to examine the longitudinal dynamics. Quite important, insights emanating from the firm and industry-based data are juxtaposed with the microdata of labour supply to understand the supply side dimensions of wage, while envisioning the implications for the economy of manufacturing to upgrade to a productivity orientation

Capturing Trends and Identifying the Emerging Cool: A Study of Indian Bollywood Celebs on Twitter

People actively participating on social networks like Facebook, Twitter and blogs are questioning the age old logic of boundaries and space. Virtual networks like twitter have given space which not only explores individual self but also connects to a mass phenomenon emerging through thought leaders, celebrities or ‘trend setters’. This study is an attempt to take the argument further and identify the emerging ‘trend’ in India through the celebtweets. To verify the argument empirically tweets of ten celebrities on twitter from Bollywood which include actors, choreographers, musicians, and producers were collected over a period of one month and tweet analysis was done by adopting hermeneutics as the method of data analysis. Appropriate codes were considered to address the major question of the paper on ‘social trends’ and the notion of ‘being cool’ to validate the question raised in the paper

Context-oriented user-centric search system for the IoT based on fuzzy clustering

The Internet of Things (IoT) paradigm envisions to support the creation of several applications that aids in the betterment of the society from various sectors such as environment, finance, industry etc. These applications are to be user-centric for their larger acceptance by the society. With the increase in the number of sensors that should are getting connected to the IoT infrastructure, there is an augmented increase in the amount of data generated by these sensors. Therefore it becomes a fundamental requirement to search for the sensors that produce the most applicable data required by the application. In this regard, context parameters of the sensors and the application users can be utilized to effectively filter out sensors from a large group. This paper proposes a sensor search scheme based on semantic-weights and fuzzy clustering. We have modified the traditional fuzzy c-means clustering algorithm by

Inhibition of IRGM establishes a robust antiviral immune state to restrict pathogenic viruses

The type I interferon (IFN) response is the major host arsenal against invading viruses. IRGM is a negative regulator of IFN responses under basal conditions. However, the role of human IRGM during viral infection has remained unclear. In this study, we show that IRGM expression is increased upon viral infection. IFN responses induced by viral PAMPs are negatively regulated by IRGM. Conversely, IRGM depletion results in a robust induction of key viral restriction factors including IFITMs, APOBECs, SAMHD1, tetherin, viperin, and HERC5/6. Additionally, antiviral processes such as MHC-I antigen presentation and stress granule signaling are enhanced in IRGM-deficient cells, indicating a robust cell-intrinsic antiviral immune state. Consistently, IRGM-depleted cells are resistant to the infection with seven viruses from five different families, including Togaviridae, Herpesviridae, Flaviviverdae, Rhabdoviridae, and Coronaviridae. Moreover, we show that Irgm1 knockout mice are highly resistant to chikungunya virus (CHIKV) infection. Altogether, our work highlights IRGM as a broad therapeutic target to promote defense against a large number of human viruses, including SARS-CoV-2, CHIKV, and Zika virus

Overexpression of the Lung Cancer-Prognostic miR-146b MicroRNAs Has a Minimal and Negative Effect on the Malignant Phenotype of A549 Lung Cancer Cells

INTRODUCTION:Expression levels of miR-146b-5p and -3p microRNAs in human non-small cell lung cancer (NSCLC) are associated with recurrence of the disease after surgery. To understand this, the effect of miR-146b overexpression was studied in A549 human lung cancer cells. METHODS:A549 cells, engineered with lentiviruses to overexpress the human pre-miR-146b precursor microRNA, were examined for proliferation, colony formation on plastic surface and in soft agar, migration and invasiveness in cell culture and in vivo in mice, chemosensitivity to cisplatin and doxorubicin, and global gene expression. miR-146b expressions were assessed in microdissected stroma and epithelia of human NSCLC tumors. Association of miR-146b-5p and -3p expression in early stage NSCLC with recurrence was analyzed. PRINCIPAL FINDINGS:A549 pre-miR-146b-overexpressors had 3-8-fold higher levels of both miR-146b microRNAs than control cells. Overexpression did not alter cellular proliferation, chemosensitivity, migration, or invasiveness; affected only 0.3% of the mRNA transcriptome; and, reduced the ability to form colonies in vitro by 25%. In human NSCLC tumors, expression of both miR-146b microRNAs was 7-10-fold higher in stroma than in cancerous epithelia, and higher miR-146b-5p but lower -3p levels were predictive of recurrence. CONCLUSIONS:Only a minimal effect of pre-miR-146b overexpression on the malignant phenotype was seen in A549 cells. This could be because of opposing effects of miR-146b-5p and -3p overexpression as suggested by the conflicting recurrence-predictive values of the two microRNAs, or because miR-146b expression changes in non-cancerous stroma and not cancerous epithelia of tumors are responsible for the prognostic value of miR-146b

Factors affecting the yield of microRNAs from laser microdissectates of formalin-fixed tissue sections

<p>Abstract</p> <p>Background</p> <p>Quantification of microRNAs in specific cell populations microdissected from tissues can be used to define their biological roles, and to develop and deploy biomarker assays. In this study, a number of variables were examined for their effect on the yield of microRNAs in samples obtained from formalin-fixed paraffin-embedded tissues by laser microdissection.</p> <p>Results</p> <p>MicroRNA yield was improved by using cresyl violet instead of hematoxylin-eosin to stain tissue sections in preparation for microdissection, silicon carbide instead of glass fiber as matrix in RNA-binding columns, and overnight digestion of dissected samples with proteinase K. Storage of slides carrying stained tissue sections at room temperature for up to a week before microdissection, and storage of the microdissectates at room temperature for up to a day before RNA extraction did not adversely affect microRNA yield.</p> <p>Conclusions</p> <p>These observations should be of value for the efficient isolation of microRNAs from microdissected formalin-fixed tissues with a flexible workflow.</p