The ability to monitor changes in the expression and localization of integrins is essential for understanding their contribution to development, tissue homeostasis and disease. Here, we pioneered the use of Crispr/Cas9 genome editing to tag an allele of the β4 subunit of the α6β4 integrin. A tdTomato tag was inserted with a linker at the COOH-terminus of β4 in mouse mammary epithelial cells. Cells harboring this tagged allele were similar to wild-type cells with respect to β4 surface expression, association with the α6 subunit, adhesion to laminin and consequent signaling. These β4 reporter cells were transformed with YAP, which enabled us to obtain novel insight into β4 dynamics in response to a migratory stimulus (scratch wound) by live-cell video microscopy. An increase in β4 expression in cells proximal to the wound edge was evident and a population of β4 expressing cells that exhibited unusually rapid migration was identified. These findings could shed insight into β4 dynamics during invasion and metastasis. Moreover, these β4 reporter cells should facilitate studies on the contribution of this integrin to mammary gland biology and cancer.
First Person is a series of interviews with the first authors of a selection of papers published in Journal of Cell Science, helping early-career researchers promote themselves alongside their papers. Ameer Elaimy is first author on ‘Real-time imaging of integrin β4 dynamics using a reporter cell line generated by Crispr/Cas9 genome editing’, published in JCS. Ameer is a MD/PhD student at University of Massachusetts Medical School in the lab of Arthur M. Mercurio, where he works on identifying mechanisms that contribute to tumorigenesis and therapy resistance in breast cancer.
Shrinkage of urban green space has led to the disruption of the ecological balance. Population growth, industrial expansion, development activities and land encroachment reduce the vegetation cover of metropolitan cities including Lahore, Pakistan. Presently, Lahore is on top of all metropolitan cities in the world having worst air quality index (AQI). Thus, monitoring of the vegetation cover in urban areas is the ultimate need of the day for the conservation and protection of environment which is also the agenda of new government (Clean and Green Pakistan). Besides, Honorable Lahore High Court has also passed an order to plant trees in the oldest and biggest graveyard of Lahore to tackle the climate change issue. Considering the above-mentioned facts, the current research was carried out for the first time in Lahore, Pakistan to analyze the vegetation cover using spatio-temporal technique. For vegetation cover assessment, spatial techniques were used in the present study viz. Normalized Difference Vegetation Index (NDVI), Transformed Normalized Vegetation Index (TNDVI), Difference Vegetation Index (DVI) and supervised classification. On the basis of high magnitude of smog and air pollution issue, four recent years were selected, i.e. 2015, 2016, 2017 and 2018 to observe ground reality. The results of the study revealed the depletion of vegetation cover in cemeteries at an alarming rate. Furthermore, the results of study revealed no significant change in green cover in Miani Sahib after the passing order of Lahore High Court of plantation in the graveyard.
Published by Oriental Scientific Publishing Company © 2019 This is an Open Access article licensed under a Creative Commons license: Attribution 4.0 International (CC-BY). Male Infertility and Viral Infection: Interference Role of the Human Herpesvirus types (3 – 6) with Disturbances Effects of Some Cytokines Hypersecretion and Seminal Oxidative Defense System in the Infertility Etiopathogenesis of Some Idiopathic Infertile Iraqi Patients
Tea waste and active carbon manufactured from dates’ stone were used as adsorbents for removal Safranin O dye and the results were compared. Safranin O dye removal efficiency was studied by batch adsorption. The experimental results displayed that the removal efficiency of dye on the two adsorbent were greatest at high acidic solution of SO solution for tea adsorbent, while for Activated Carbon at high alkalinity for the same dye. This study shows the adsorption equilibrium isotherms were followed Freundlich and Langmuir isotherms, where safranin O dye adsorption obey Freundlich isotherm. The effect of temperature was studied and thermodynamic parameters were calculated.
The ReCap Femoral Resurfacing System has been associated with increased cases of revision surgery when compared to other hip resurfacing systems. However, computer-assisted navigation may have the potential to reduce the risk of post-operative complications by providing more accurate intraoperative measurements for acetabular component positioning.
Disconnected interacting protein 2 homolog A (DIP2A) is highly expressed in nervous system and respiratory system of developing embryos. However, genes regulated by Dip2a in developing brain and lung have not been systematically studied. Transcriptome of brain and lung in embryonic 19.5 day (E19.5) were compared between wild type and Dip2a -/- mice. An average of 50 million reads per sample was mapped to the reference sequence. A total of 214 DEGs were detected in brain (82 up and 132 down) and 1900 DEGs in lung (1259 up and 641 down). GO enrichment analysis indicated that DEGs in both Brain and Lung were mainly enriched in biological processes ‘DNA-templated transcription and Transcription from RNA polymerase II promoter’, ‘multicellular organism development’, ‘cell differentiation’ and ‘apoptotic process’. In addition, COG classification showed that both were mostly involved in ‘Replication, Recombination, and Repair’, ‘Signal transduction and mechanism’, ‘Translation, Ribosomal structure and Biogenesis’ and ‘Transcription’. KEGG enrichment analysis showed that brain was mainly enriched in ‘Thyroid cancer’ pathway whereas lung in ‘Complement and Coagulation Cascades’ pathway. Transcription factor (TF) annotation analysis identified Zinc finger domain containing (ZF) proteins were mostly regulated in lung and brain. Interestingly, study identified genes Skor2 , Gpr3711 , Runx1 , Erbb3 , Frmd7 , Fut10 , Sox11 , Hapln1 , Tfap2c and Plxnb3 from brain that play important roles in neuronal cell maturation, differentiation, and survival; genes Hoxa5 , Eya1 , Errfi1 , Sox11 , Shh , Igf1 , Ccbe1 , Crh , Fgf9 , Lama5 , Pdgfra , Ptn , Rbp4 and Wnt7a from lung are important in lung development. Expression levels of the candidate genes were validated by qRT-PCR. Genome wide transcriptional analysis using wild type and Dip2a knockout mice in brain and lung at embryonic day 19.5 (E19.5) provided a genetic basis of molecular function of these genes.
Background Cardiovascular diseases (CVD) are rising in India resulting in major health system challenges. Methods Eighteen primary health centre (PHC) clusters in rural Andhra Pradesh were randomised over three, 6-month steps to an intervention comprising: (1) household CVD risk assessments by village-based community health workers (CHWs) using a mobile tablet device; (2) electronic referral and clinical decision support for PHC doctors; and (3) a tracking system for follow-up care. Independent data collectors screened people aged ≥ 40 years in 54 villages serviced by the PHCs to create a high CVD risk cohort (based on WHO risk charts and blood pressure thresholds). Randomly selected, independent samples, comprising 15% of this cohort, were reviewed at each 6-month step. The primary outcome was the proportion meeting systolic blood pressure (SBP) targets (<140mmHg). Findings Eight-four percent of the eligible population (n = 62,254) were assessed at baseline (18.4% at high CVD risk). Of those at high risk, 75.3% were followed up over two years. CHWs screened 85.9% of the baseline cohort and doctors followed up 70.0% of all high risk referrals. There was no difference in the proportion of people achieving SBP targets (41.2% vs 39.2%; adjusted odds ratio (OR) 1.01 95% CI 0.76–1.35) or receiving BP-lowering medications in the intervention vs control periods respectively. There was a high discordance in risk scores generated by independent data collectors and CHWs, resulting in only 37.2% of the evaluation cohort exposed to the intervention. This discordance was mainly driven by fluctuating BP values (both normal variability and marked seasonal variations). In the pre-specified high risk concordant subgroup, there was greater use of BP-lowering medications in the intervention period (54.3% vs 47.9%, OR 1.22, 95% CI 1.03–1.44) but no impact on BP control. Conclusions The strategy was well implemented with increased treatment rates among high risk individuals assessed by CHWs, however effects on BP were not demonstrated. Use of guideline-recommended BP thresholds for identifying high risk individuals substantially affected the reproducibility of risk assessment, and thus the ability to reliably evaluate the effectiveness of the intervention. In addition, unanticipated seasonal variation in BP in the context of a stepped-wedge trial highlights the inherent risks of this study design. Trial registration Clinical Trials Registry of India CTRI/2013/06/ 003753.
Providing the dairy industry with an effective and safe disinfectant is considered a key step in improving the farm hygiene and biosecurity. Salmonella infection via foodborne transmission remains a major public health threat. The main objective of this study was therefore to characterize and compare the decontamination power of NaOCl, aqueous-O 3 , and PAA against cattle manure based- Salmonella heavily contaminated various surfaces (plastic, nylon, rubber, and wood) using Bayesian analysis. In a crossover design, 14 strips of each material were randomly assigned between 3 groups, treatment (n = 6), positive-control (contaminated with feces- Salmonella mixture, but not exposed to disinfectants, n = 6), and negative control (laboratory blank, inoculated only with sterile water, n = 2). The strips were soaked in cattle manure inoculated with 10 7 –10 8 of Salmonella Typhimurium-Choleraesuis (aSTC) and exposed to 50 mL of 200 ppm NaOCl, 9 ppm aqueous-O 3 , or 400 ppm PAA for 4 minutes. Bayesian methods were used for analysis. On plastic and nylon surfaces, NaOCl, aqueous-O 3, or PAA reduce aSTC population to a safe level (>5.0-log 10 ) within 4 minutes. On rubber surface, PAA and aqueous-O 3 can produce a reduction in aSTC population 50% and 30% higher than NaOCl with posterior probabilities of 97% and 90%, respectively. However, PAA can produce reduction factor on wood surface 40% higher than aqueous-O 3 and NaOCl with posterior probabilities of 97% and 73%, respectively. We conclude that smooth surfaces were most effectively decontaminated. Peracetic acid of 400 ppm can provide an effective means for controlling Salmonella population heavily contaminated various surfaces in dairy operations. However, the safe residues and strong reactivity makes aqueous-O 3 and PAA attractive alternative disinfectants for improving farm hygiene and biosecurity.