Characterization of a “hypothetical protein”, EF1025, from Enterococcus faecalis: role in cell length and shape

DivIVA plays multifaceted roles in Gram-positive organisms by associating with various cell division and non-cell division proteins. While the interaction of DivIVA with other proteins has been studied in many Gram-positive bacteria, no information is available about DivIVA- associating proteins in E. faecalis. This research reports a novel DivIVAEf interacting protein named EF1025 (encoded by EF1025) (confirmed using Bacterial Two-Hybrid, Glutathione S-Transferase pull-down, and co-immunoprecipitation assays) that affects cell length and morphology in E. faecalis. EF1025 is predominantly conserved in Gram-positive bacteria and contains a conserved N-terminal DNA binding Helix-turn-Helix (HTH) domain and two Cystathionine β-Synthase (CBS) domains located centrally and at the C-terminus. The protein, EF1025, oligomerizes to form a higher-order oligomer and the two CBS domains are responsible for its self-interaction. Viable cells were recovered after insertional inactivation or deletion of EF1025 only through complementation of EF1025 in trans. These cells were longer than the average length of E. faecalis cells and had distorted shapes. Overexpression of EF1025 also resulted in cell elongation but had no effect on cell shape. Immuno-staining revealed comparable localization patterns of EF1025 and DivIVAEf in the later stages of division in E. faecalis cells. The EF1025 homologue in Bacillus subtilis, CcpN, is a transcriptional repressor in Bacillus subtilis. In the presence of glucose, CcpN binds to the promoter region of gapB and pckA and downregulates their expression. CcpN interacted with DivIVA of B. subtilis in B2H and GST-pull down assays. A heterologous interaction between EF1025 and DivIVABs was also identified in a GST-pull down assay. Insertional inactivation of ccpN leads to cell elongation and growth of cells in straight chains. These findings suggest an additional function of CcpN in B. subtilis, therefore, CcpN is a dual function performing protein involved in both gluconeogenesis and cell elongation. E. faecalis contains homologues of divisome proteins FtsZ, FtsA, FtsK, FtsQ, FtsL, FtsI and FtsB, however, the cell division interactome of E. faecalis, by contrast, is not presently known. This thesis also presents the unique interactome of E. faecalis divisome proteins (i.e. FtsZEf, FtsAEf, FtsQEf, FtsLEf, FtsIEf, FtsWEf, DivIVAEf, and FtsBEf), established using Bacterial-two hybrid system. The interaction of FtsA with FtsI, FtsL, and FtsZ, is common among E. faecalis, S. pneumoniae and S. aureus cell division interactomes. One unique interaction i.e. FtsZEf-FtsIEf was identified in E. faecalis cell division interactome. While studying the divisome interactome of E. faecalis, it was observed that EF1025 is not a part of the divisome machinery in E. faecalis as it did not interact with any divisome protein except DivIVAEf

Are we assessing them right? A study on the content validity of undergraduate pathology question papers and role of blueprinting in assessment

Background: Assessment forms a very essential and critical part of the learning process in medical education. Written examinations are widely used to test the knowledge component of learning. For a written examination to be valid, it should match the contents of the course and should provide proportional weightage to each of the content. A periodic evaluation of question papers to examine the weightage of each topic and the use of blue printing is necessary to ensure the validity of the written assessments.Methods: The ten year question papers (2007-2016) of II MBBS summative written examinations in Pathology conducted by Rajasthan University of Health Sciences (RUHS) were analysed and the appropriateness of weightage given to content areas was examined in relation to the syllabus.Results: The syllabus distribution in Paper 1 and Paper 2 was highly skewed with extreme concentration of topics in Paper 1 (due to clubbing of both general and systemic pathology) which led to under-representation of many topics despite their high Impact and Frequency. There was also evidence of paper setter’s bias leading to disproportionate (over/under) representation of many topics. An ideal blueprint for paper 1 and 2 was prepared with appropriate syllabus distribution and allotment of marks as per weightage of each topic according to its impact and frequency score.Conclusions: Frequent analysis should be carried out to provide feedback and to ensure that the assessments are aligned with the learning objectives. A blueprint is a vital component and helps us to plan written assessments in a rational and balanced manner

Negotiating conflicting discourses of quality teaching in Fiji: initial teacher education and practicum at the University of the South Pacific

This article identifies a number of conflicting discourses informing education in Fiji and their impact on Initial Teacher Education (ITE) students. The socially constructivist progressivism of the Ministry of Education and the ITE provider is being eroded by a set of socially conservative discourses symptomatic of neoliberal education reforms elsewhere. It is the Practicum where the conflict is most acutely evidenced. To highlight the conflict 90 ITE students, as ethno-graphic fieldworkers, have used an accepted quality teaching checklist to record the teaching they witnessed while on practicum. The resulting misalignments between discourses of quality teaching identified in this article and highlighted by ITE students contributes to debates about what constitutes effective teaching in Fiji. Additionally, despite the multi-discursive reality of Fijian education the article suggests ITE based on a learning-centred rather than learner-centred approach where teachers make critical choices for teaching based on links between pedagogy, context and consequence

Evaluation of High Performance Converters Under Low Dose Rate Total Ionizing Dose (TID) Testing for NASA Programs

This paper reports the results of low dose rate (0.01-0.18 rads(Si)/sec) total ionizing dose (TID) tests performed on several types of high performance converters. The parts used in this evaluation represented devices such as a high speed flash converter, a 16-bit ADC and a voltage-to-frequency converter

Evaluation of interleukin-10 production in Pseudomonas aeruginosa induced acute pyelonephritis

SummaryPseudomonas aeruginosa is an opportunistic pathogen of immunocompromised hosts. This pathogen has a tendency to form biofilms on the surface of indwelling catheters leading to acute and chronic urinary tract infections that result in significant morbidity and mortality. In the present study, kinetics of interleukin-10 (IL-10) production in mouse renal tissue was studied employing experimental mouse model of acute pyelonephritis induced with planktonic and biofilm cells of P. aeruginosa. IL-10 production was found to be significantly lower in biofilm cell instilled mice compared to planktonic cell infected animals, which corroborated with higher bacterial load and tissue damage. The data suggests that downregulation of IL-10 production may be novel strategy employed by biofilm cells to cause tissue damage and hence bacterial persistence. The results of the present study may open up avenues of research that will ultimately provide the foundation for the development of preventative measures and therapeutic strategies to successfully treat P. aeruginosa biofilm infections based on the administration of anti-inflammatory agents

Coronal pulp cavity index as noble modality for age estimation : a digital image analysis

Funding Information: The financial support provided by UGC, New Delhi, in the form of “UGC-JRF Fellowship” (Letter No F.15-9 (JUNE 2014)/2014(NET), helped the author in preparation of this research article.Peer reviewe

Analysis of caesarean section rate according to modified Robson’s classification at tertiary care centre in Uttarakhand, India

Background: High caesarean section rate worldwide including India is matter of concern. The aim of this study is to analyse caesarean section rate at tertiary care centre according to Modified Robson’s classification.Methods: This retrospective study was conducted at Shri Guru Ram Rai Institute of Medical and Health Sciences (SGRRIMHS) and Shri Mahant Indiresh Hospital at Dehradun from April 2018 to September 2018. All women delivered during this period were classified according to modified Robson’s classification using their maternal characteristics and obstetric history. For each group, authors calculated the caesarean section rate within the group and its contribution to overall caesarean section rate.Results: Out of total 1302 women delivered, 395 underwent CS (30.3%).The major contribution to overall caesarean section rate was 33.4% by group 5 (Previous CS, singleton, cephalic, >37weeks) followed by 16.7% by group 1 (nullipara, singleton, cephalic, >37 weeks, spontaneous labour), 12.4% by group 3 ( multipara, singleton, cephalic, >37 weeks, spontaneous labour ).CS rates among various group ranges from 100% among women with abnormal lie (group 9) to 77.5% in nulliparous breech (group 6), 73.7% in previous CS (group 5) and least 11.2%  in multipara induced or pre labour CS (group 4).Conclusions: Modified Robson classification is simple, systematic, reproducible and can be effectively utilized in analyzing delivering women. Major contribution to overall caesarean section is made by previous CS

Study of image fusion using discrete wavelet and multiwavelet transform

ABSTRACT: Image fusion is processes of combining complementary information from a set of input images. The resultant fused image give large and reliable information. In this paper we study about Discrete wavelet and Discrete Multiwavelet and there use in image fusion. Discrete wavelet transform (DWT) technique is used for multi Resolution fusion. Multi Resolution fusion uses wavelet transform at multi scale for the representation of the source images. Multiwavelets are extension of scalar wavelets, and have many advantages over scalar wavelets. Multiwavelet analysis can provide a more absolute image analysis than wavelet multiresolution analysis.In this paper DWT and DMWT are qualitatively compaired with each other Keywords: Image fusion, multiwavelet transform,wavelet transform . multisensor image . I.INTRODUCTION Image Fusion is defined as the task or technique of combining two or more images into a single image. The new single image retains important information from each input image. Image fusion is a powerful tool used to increase the quality of image. Image fusion increases reliability, decreases uncertainty and storage cost by a single informative image than storing multiple images Image fusion can take place at three different levels pixel feature,decision level Image fusion technique can be classified into two categories -Direct Image Fusion and Multi resolution Image Fusion. Multiresolution Image fusion techniques based on pixel level fusion methods. Multi Resolution fusion uses wavelet and multiwavelet transform at multi scale for the representation of the input images. Image fusion based on the DWT can provide better performance than fusion based on other multiscale methods such as Laplacian pyramid, morphological pyramid. Wavelet transform in multiresolution can provide good localization in both frequency and space domains. In comparision with other multiscale transforms, the discretewavelet transform is more compact, and give detail about directional information in the low-low, high-low, low-high, and high-high bands, and contains unique information at different resolutions. The main dwarback of the scalar wavelet functions is time-frequency localization property. Multiwavelets have more than two scaling and wavelet functions. The multi-wavelet has many outstanding properties like orthogonality, short support, symmetry, and high degree of vanishing moments which is desirable for image processing . A multiwavelet system provides perfect reconstruction also preserve length (orthogonality), good performance at the boundaries , and a high order of approximation . By this multiwavelets gives superior performance for image processing applications as compared with the scalar wavelets. II.IMAGE FUSION: SCALAR WAVELETS The wavelet transform is use to detect local features in a signal process. It also used for decomposition of two dimensional (2D) signals such as 2D gray-scale image signals for multiresolution analysis. In wavelet transforms a signal is decomposed in lower frequency band and high frequency bands. In discrete wavelet transform (DWT), twochannel filter bank is used. When decomposition is performed, the approximation and detail component can be separated 2-D Discrete Wavelet Transformation (DWT) converts the image from the spatial domain to frequency domain. The image is divided by vertical and horizontal lines and represents the first-order of DWT, and the image can be separated with four parts those are LL1, LH1, HL1 and HH1. [10