Use of integrase-minus lentiviral vector for transient expression

Objective: Lentivirus-derived vectors are among the most promising viral vectors for gene therapy which is currently available, but their use in clinical practice is limited due to associated risk of insertional mutagenesis. Gene targeting is an ideal method for gene therapy, but it has low efficiency in comparison to viral vector methods. In this study, we are going to design and construct an integrase-minus lentiviral vector. This vector is suitable for transient expression of gene and gene targeting with viral vector. Materials and Methods: In this experimental study, three missense mutations were induced in the catalytic domain of Integrase gene in the pLP1 plasmid and resulted D64V, D116A and E152G changes in the amino acid sequence through site directed mutagenesis. The pLenti6.2-GW/EmGFP transfer vector, associated with native and mutated packaging mix, was transfected into 293T cell line. In order to titer the lentivirus stock, the viruses were harvested. Finally, the viruses transduced into COS-7 cell line to assess green fluorescent protein (GFP) gene expression by a fluorescence microscopy. Results: Recombinant and wild lentiviruses titer was about 5�8�10 6 transducing units/ ml in COS-7 cell line. The number of GFP-positive cells transduced with native viruses was decreased slightly during two weeks after viral transduction. In contrast, in the case of integrase-minus viruses, a dramatic decrease in the number of GFP positive cells was observed. Conclusion: This study was conducted to overcome the integration of lentiviral genome into a host genome. Nonintegrating lentiviral vectors can be used for transient gene expression and gene targeting if a Target gene cassette is placed in the lentivirus gene structure. This combination method decreases disadvantages of both processes, such as random integration of lentiviruses and low efficiency of gene targeting

An Experimental-Intelligent Method to Predict Noise Value of Drilling in Dimension Stone Industry

The noise of drilling in the dimension stone business is unbearable for both the workplace and the people who work there. In order to reduce the negative effects drilling has on the health of the environment, the drilling noise has to be measured, assessed, and controlled. The main purpose of this work is to investigate an experimental-intelligent method to predict the noise value of drilling in the dimension stone industry. For this purpose, 135 laboratory tests are designed on five types of rocks (four types of hard rock and one type of soft rock): and their results are measured in the first step. In the second step, due to the unpredicted and uncertain issues in this case, artificial intelligence (AI) approaches are applied, and the modeling is conducted using three intelligent systems (IS): namely an adaptive neuro-fuzzy inference system-SCM (ANFIS-SCM): an adaptive neuro-fuzzy inference system-FCM (ANFIS-FCM): and the radial basis function network (RBF) neural network. 75% of the samples are considered for training, and the rest for testing. Several models are constructed, and the results indicate that although there is no significant difference between the models according to the performance indices, the proposed construction of ANFIS-SCM can be considered as an efficient tool in the evaluation of drilling noise. Finally, several scenarios are designed with different input modes, and the results obtained prove that the types of rock and the drill bits are more important than the operational characteristics of the machine

Estimation of topographical effects at Narni ridge (Central Italy): comparisons between experimental results and numerical modelling

In the present work the seismic site response of Narni ridge (central Italy) is evaluated by comparing experimental results and numerical simulations. The inhabited village of Narni is located in the central Italian Apennines at the top of a steep massive limestone ridge. From March to September 2009 the site was instrumented with 10 weak-motion stations, 3 of which located at the base of the ridge and 7 at the top. The velocimetric network recorded 642 events of ML up to 5.3 and hypocentral distance up to about 100 km. The great amount of data are related to the April 2009 L’Aquila sequence. The site response was analyzed using both reference (SSR, Standard Spectral Ratio) and non reference spectral techniques (HVSR, Horizontal to Vertical Spectral Ratio). Moreover directional analyses were performed in order to evaluate the influence of the ridge orientation with respect to the selected source-site paths. In general the experimental results show amplification factors for frequencies between 4 and 5 Hz for almost all stations installed along the crest. The SSR technique provides amplification factors up to 4.5 detected considering directions perpendicular to the main elongation of the ridge. The results obtained from the monitoring activity were used as a target for bidimensional and tridimensional numerical simulations, performed using a hybrid finite-boundary element method for 2D and a boundary element method for 3D analyses respectively. In general, the results obtained through numerical simulation fit well the experimental data in terms of range of amplified frequencies, but they underestimate by a factor of about 2 the related amplification factors with respect to the observations

Land subsidence hazard in iran revealed by country-scale analysis of sentinel-1 insar

Many areas across Iran are subject to land subsidence, a sign of exceeding stress due to the over-extraction of groundwater during the past decades. This paper uses a huge dataset of Sentinel-1, acquired since 2014 in 66 image frames of 250×250km, to identify and monitor land subsidence across Iran. Using a two-step time series analysis, we first identify subsidence zones at a medium scale of 100m across the country. For the first time, our results provide a comprehensive nationwide map of subsidence in Iran and recognize its spatial distribution and magnitude. Then, in the second step of analysis, we quantify the deformation time series at the highest possible resolution to study its impact on civil infrastructure. The results spots the hazard posed by land subsidence to different infrastructure. Examples of road and railways affected by land subsidence hazard in Tehran and Mashhad, two of the most populated cities in Iran, are presented in this study

Exploring cloud-based platforms for rapid insar time series analysis

The idea of near real-time deformation analysis using Synthetic Aperture Radar (SAR) data as a response to natural and anthropogenic disasters has been an interesting topic in the last years. A major limiting factor for this purpose has been the non-availability of both spatially and temporally homogeneous SAR datasets. This has now been resolved thanks to the SAR data provided by the Sentinel-1A/B missions, freely available at a global scale via the Copernicus program of the European Space Agency (ESA). Efficient InSAR analysis in the era of Sentinel demands working with cloud-based platforms to tackle problems posed by large volumes of data. In this study, we explore a variety of existing cloud-based platforms for Multioral Interferometric SAR (MTI) analysis and discuss their opportunities and limitations

Tychiini and Mecinini (Coleoptera: Curculionidae, Curculioninae) of Iran: eleven species recorded for the first time, with new data on host plants and distribution of several species

A faunistic study on the tribes Tychiini and Mecinini (Curculionidae, Curculioninae) was carried out during the years 2010-2013 in different ecological regions of Iran. Twenty nine species belonging to the genera Mecinus Germar, 1821, Gymnetron Schoenherr, 1825, Rhinusa Stephens, 1829, Cleopomiarus Pierce, 1919, Tychius Germar, 1817 and Sibinia Germar, 1817 were collected. Localities and ecological notes on each species are provided. Among these, The following 11 species are new to the Iranian fauna: Mecinus crassifemur (Arzanov, 1991), Mecinus simus (Mulsant & Rey, 1859), Gymnetron linkei Reitter, 1907, Rhinusa antirrhini (Paykull, 1800), Rhinusa brondelii (H. Brisout de Barneville, 1862), Rhinusa florum (Rübsaamen, 1895), Tychius reitteri Faust, 1889, Tychius tridentinus Penecke, 1922, Sibinia aureofulva (Desbrochers des Loges, 1875), Sibinia pellucens (Scopoli, 1772), Sibinia unicolor Fảhraeus, 1843, whereas all other 18 species are new for one or more provinces. Host plants of several species reported for the first time and commented as well

Clinical Utility of a Unique Genome-Wide DNA Methylation Signature for KMT2A-Related Syndrome

Wiedemann\u2013Steiner syndrome (WDSTS) is a Mendelian syndromic intellectual disability (ID) condition associated with hypertrichosis cubiti, short stature, and characteristic facies caused by pathogenic variants in the KMT2A gene. Clinical features can be inconclusive in mild and unusual WDSTS presentations with variable ID (mild to severe), facies (typical or not) and other associated malformations (bone, cerebral, renal, cardiac and ophthalmological anomalies). Interpretation and classification of rare KMT2A variants can be challenging. A genome-wide DNA methylation episignature for KMT2A-related syndrome could allow functional classification of variants and provide insights into the pathophysiology of WDSTS. Therefore, we assessed genome-wide DNA methylation profiles in a cohort of 60 patients with clinical diagnosis for WDSTS or Kabuki and identified a unique highly sensitive and specific DNA methylation episignature as a molecular biomarker of WDSTS. WDSTS episignature enabled classification of variants of uncertain significance in the KMT2A gene as well as confirmation of diagnosis in patients with clinical presentation of WDSTS without known genetic variants. The changes in the methylation profile resulting from KMT2A mutations involve global reduction in methylation in various genes, including homeobox gene promoters. These findings provide novel insights into the molecular etiology of WDSTS and explain the broad phenotypic spectrum of the disease

Association of high risk human papillomavirus and breast cancer : a UK based study

Infection by human papillomaviruses (HPVs) has been implicated in the aetiology of a variety of cancers. Studies evaluating the presence of HPVs in breast cancer (BC) have generated considerable controversy. To date, most studies have focused on the presence of viral DNA in BC; however there are important gaps in evidencing the role of HPV persistence in the invasiveness of BC. While these studies have been conducted in several countries, none, on the presence and biological activity of high risk (HR) HPV in BC has been done in the UK. Hence, we aimed to investigate these gaps by screening a total of 110 fresh breast tissue specimens from UK patients for the presence of twelve HR-HPV types DNA using PCR and Sanger sequencing. Samples positive for HPV-DNA were screened for viral oncoprotein expression using western blot and dot blot. Data obtained showed the presence of HR-HPVs in 42% of breast tissues of which the viral activity was only confirmed in a number of invasive carcinomas (5/26). This finding, the first to report in the UK, suggests that the selective expression of viral oncoprotein in invasive cases may propose a role for HR-HPVs in the development of some types of BC