Stable Knockdown of Adenosine Kinase by Lentiviral Anti-ADK miR-shRNAs in Wharton�s Jelly Stem Cells

Objective: In this study, we describe an efficient approach for stable knockdown of adenosine kinase (ADK) using lentiviral system, in an astrocytoma cell line and in human Wharton�s jelly mesenchymal stem cells (hWJMSCs). These sources of stem cells besides having multilineage differentiation potential and immunomodulatory activities, are easily available in unlimited numbers, do not raise ethical concerns and are attractive for gene manipulation and cell-based gene therapy. Materials and Methods: In this experimental study, we targeted adenosine kinase mRNA at 3' and performed coding sequences using eight miR-based expressing cassettes of anti-ADK short hairpin RNA (shRNAs). First, these cassettes with scrambled control sequences were cloned into expressing lentiviral pGIPZ vector. Quantitative real time-polymerase chain reaction (qRT-PCR) was used to screen multi-cassettes anti-ADK miR-shRNAs in stably transduced U-251 MG cell line and measuring ADK gene expression at mRNA level. Extracted WJMSCs were characterized using flow cytometry for expressing mesenchymal specific marker (CD44+) and lack of expression of hematopoietic lineage marker (CD45-). Then, the lentiviral vector that expressed the most efficient anti-ADK miR-shRNA, was employed to stably transduce WJMSCs. Results: Transfection of anti-ADK miR-shRNAs in HEK293T cells using CaPO4 method showed high efficiency. We successfully transduced U-251 cell line by recombinant lentiviruses and screened eight cassettes of anti-ADK miR-shRNAs in stably transduced U-251 MG cell line by qRT-PCR. RNAi-mediated down-regulation of ADK by lentiviral system indicated up to 95 down-regulation of ADK. Following lentiviral transduction of WJMSCs with anti-ADK miR-shRNA expression cassette, we also implicated, down-regulation of ADK up to 95 by qRT-PCR and confirmed it by western blot analysis at the protein level. Conclusion: Our findings indicate efficient usage of shRNA cassette for ADK knockdown. Engineered WJMSCs with genome editing methods like CRISPR/cas9 or more safe viral systems such as adeno-associated vectors (AAV) might be an attractive source in cell-based gene therapy and may have therapeutic potential for epilepsy

Stable Knockdown of Adenosine Kinase by Lentiviral Anti-ADK miR-shRNAs in Wharton�s Jelly Stem Cells

Objective: In this study, we describe an efficient approach for stable knockdown of adenosine kinase (ADK) using lentiviral system, in an astrocytoma cell line and in human Wharton�s jelly mesenchymal stem cells (hWJMSCs). These sources of stem cells besides having multilineage differentiation potential and immunomodulatory activities, are easily available in unlimited numbers, do not raise ethical concerns and are attractive for gene manipulation and cell-based gene therapy. Materials and Methods: In this experimental study, we targeted adenosine kinase mRNA at 3' and performed coding sequences using eight miR-based expressing cassettes of anti-ADK short hairpin RNA (shRNAs). First, these cassettes with scrambled control sequences were cloned into expressing lentiviral pGIPZ vector. Quantitative real time-polymerase chain reaction (qRT-PCR) was used to screen multi-cassettes anti-ADK miR-shRNAs in stably transduced U-251 MG cell line and measuring ADK gene expression at mRNA level. Extracted WJMSCs were characterized using flow cytometry for expressing mesenchymal specific marker (CD44+) and lack of expression of hematopoietic lineage marker (CD45-). Then, the lentiviral vector that expressed the most efficient anti-ADK miR-shRNA, was employed to stably transduce WJMSCs. Results: Transfection of anti-ADK miR-shRNAs in HEK293T cells using CaPO4 method showed high efficiency. We successfully transduced U-251 cell line by recombinant lentiviruses and screened eight cassettes of anti-ADK miR-shRNAs in stably transduced U-251 MG cell line by qRT-PCR. RNAi-mediated down-regulation of ADK by lentiviral system indicated up to 95 down-regulation of ADK. Following lentiviral transduction of WJMSCs with anti-ADK miR-shRNA expression cassette, we also implicated, down-regulation of ADK up to 95 by qRT-PCR and confirmed it by western blot analysis at the protein level. Conclusion: Our findings indicate efficient usage of shRNA cassette for ADK knockdown. Engineered WJMSCs with genome editing methods like CRISPR/cas9 or more safe viral systems such as adeno-associated vectors (AAV) might be an attractive source in cell-based gene therapy and may have therapeutic potential for epilepsy

Does technology and Innovation Management improve Market Position? Empirical Evidence from Innovating Firms in South Africa

There is a growing recognition of the central role of technology and knowledge management for market success of organizations. Little is empirically know, however, about this relationship. Drawing on the South African Innovation Survey, a unique dataset on innovative behavior of South African firms in manufacturing and services, this paper investigates the question to what extent and in which ways do technology and innovation management activities affect firms’ market position. Findings show that conducting technology strategy activities pays out. Moreover, especially a combination of internal and external technology audits seems to be beneficial for organizational performance

Observation of inverse Compton emission from a long γ-ray burst.

Long-duration γ-ray bursts (GRBs) originate from ultra-relativistic jets launched from the collapsing cores of dying massive stars. They are characterized by an initial phase of bright and highly variable radiation in the kiloelectronvolt-to-megaelectronvolt band, which is probably produced within the jet and lasts from milliseconds to minutes, known as the prompt emission1,2. Subsequently, the interaction of the jet with the surrounding medium generates shock waves that are responsible for the afterglow emission, which lasts from days to months and occurs over a broad energy range from the radio to the gigaelectronvolt bands1-6. The afterglow emission is generally well explained as synchrotron radiation emitted by electrons accelerated by the external shock7-9. Recently, intense long-lasting emission between 0.2 and 1 teraelectronvolts was observed from GRB 190114C10,11. Here we report multi-frequency observations of GRB 190114C, and study the evolution in time of the GRB emission across 17 orders of magnitude in energy, from 5 × 10-6 to 1012 electronvolts. We find that the broadband spectral energy distribution is double-peaked, with the teraelectronvolt emission constituting a distinct spectral component with power comparable to the synchrotron component. This component is associated with the afterglow and is satisfactorily explained by inverse Compton up-scattering of synchrotron photons by high-energy electrons. We find that the conditions required to account for the observed teraelectronvolt component are typical for GRBs, supporting the possibility that inverse Compton emission is commonly produced in GRBs

Viewpoints of students of Shahid Sadoughi University of Medical Sciences regarding their academic advisors

Introduction: Academic advisors have been proven important in students’ success. Insufficient guidance and counseling can delay professional and educational achievements. Proper counseling can provide opportunities for educational success and also positive change students’ behavioral models. This study was accomplished with the purpose of assigning the function of advisors from the viewpoint of the students. Methods: This descriptive study was done on 385 students of Shahid Sadoughi University. Randomized stratified sampling was used. The data were collected by completing the self-reported questionnaire and then were analyzed by SPSS software. Advisors’ functions in each of the items were evaluated. The significance level of 0.05 was considered. Results: The majority of samples were from Nursing, Midwifery, and Public Health School and a few from Faculty of Medicine. The majority of students (41.8%) agreed to have the same advisor from the first term to the last. Also the results showed that the functions of advisors were poor in the areas of constant supervision and monitoring of students’ educational status (35.8%), preparing the timetable for performing the personal or group counseling (41.8%) and providing counseling sessions according to the timetable (42.3%). But their functions were fair in the areas of acceptable guidance of students in the educational problems (35.1%), and the advisor’s familiarity to educational regulations (43.9%). It is necessary to note that the advisor's ability in making the intimate relationship with students (37.1%), and the advisors’ motivation and tendency to guidance and counseling (30.1%) were the two items evaluated excellent by students. Conclusion: It is necessary that advisors make the students familiar with educational regulations at the first semester of higher education. Also, the policies and regulations for advisors will clarify their activities and will help them to perform better

Incidence of low birth weight and associated risk factors during March 2002-2003 in Tonekabon, Iran

Background and purpose: Low birth weight (LBW; birthweight 2500 g. or below) is a public health problem, because it is associated with increased risk of morbidity and mortality. This prospective study was conducted during March 2002 - 2003, to determine the incidence of low birth weight infants and associated risk factors in neonates born at Shahid Rajaee Hospital in Tonekabone, Iran.Materials and methods: For every LBW case, maternal age, sex, gestational age, parity, route of delivery and spacing between pregnancies of low birth weight were analyzed. For control, the first baby born before and immediately after the LBW baby who weighed more than 2500 g. at birth were analyzed.Results: The results revealed that the incidence of low birth weight was 4.2 % which is lower than population of other cities and only 0.34% (7 cases) of newborns weighted 1500 g. or less. The higher incidence of low birth weight was found in the primigravid (P=0.042), in cesarean section (P=0.025) and close spacing of pregnancy (P=0.033). Maternal age and sex of baby had no significant effect on the incidence of low birth weight. There were more premature deliveries in cases than controls (P=0.000).Conclusion: According to present findings, LBW incidence of albeit in Tonekabon in comparison with other cities throught the country is at acceptable level, prevention of premature delivery and conduction of health education programs for high risk groups, especially primigravid can lower the rates