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

Two novel splice variants of SOX2OT, SOX2OT-S1, and SOX2OT-S2 are coupregulated with SOX2 and OCT4 in esophageal squamous cell carcinoma

Long noncoding RNAs (lncRNAs) have emerged as new regulators of stem cell pluripotency and tumorigenesis. The SOX2 gene, a master regulator of pluripotency, is embedded within the third intron of a lncRNA known as SOX2 overlapping transcript (SOX2OT). SOX2OT has been suspected to participate in regulation of SOX2 expression and/or other related processes; nevertheless, its potential involvement in tumor initiation and/or progression is unclear. Here, we have evaluated a possible correlation between expression patterns of SOX2OT and those of master regulators of pluripotency, SOX2 and OCT4, in esophageal squamous cell carcinoma (ESCC) tissue samples. We have also examined its potential function in the human embryonic carcinoma stem cell line, NTERA2 (NT2), which highly expresses SOX2OT, SOX2, and OCT4. Our data revealed a significant coupregulation of SOX2OT along with SOX2 and OCT4 in tumor samples, compared to the non-tumor tissues obtained from the margin of same tumors. We also identified two novel splice variants of SOX2OT (SOX2OT-S1 and SOX2OT-S2) which coupregulated with SOX2 and OCT4 in ESCCs. Suppressing SOX2OT variants caused a profound alteration in cell cycle distribution, including a 5.9 and 6.9 time increase in sub-G1 phase of cell cycle for SOX2OT-S1 and SOX2OT-S2, respectively. The expression of all variants was significantly diminished, upon the induction of neural differentiation in NT2 cells, suggesting their potential functional links to the undifferentiated state of the cells. Our data suggest a part for SOX2OT spliced variants in tumor initiation and/or progression as well as regulating pluripotent state of stem cells. © AlphaMed Press 2013

Numerical investigation on the position of holes for reducing stress concentration in composite plates with bolted and riveted joints

AbstractThis paper studies the effects of fiber orientaion and holes position on stress concentration and the determination of weakened areas in the composite of glass fiber reinforced epoxy resin around the hole for joints by using the finite element method. In this study, for the observation of areas affected by stress concentration Tsai-Wu failure criterion is used to determine the failed elements and ANSYS Software is implemented for modeling. In order to compare the effect of geometric parameters on stress concentration around the holes, two types of hole position arrangement along with fibers orientation have been studied. Results show that the stress concentration coefficient is lower in the second type of holes arrangement in comparison with the first type for the same component dimensions. Increasing the distance from hole center to upper or lower edge of the sample and also decreasing the distance between holes, would result in an increase in the stress concentration

Isolation and Kinetic Modeling of New Culture from Compost with High Capability of Degrading n-Hexadecane, Focused on Ochrobactrum Oryzae and Paenibacillus Lautus

Nowadays, petroleum pollution is one of the most important environmental challenges in Iran. Some bacteria were isolated for hexadecane degrading from soil, sediment, and sludge; however, there is no report on its isolation from compost. This study was aimed to isolation, molecular identification of novel bacteria with high capability of hexadecane-degrading from compost using enrichments media. The isolated bacteria were identified by PCR with 16S rDNA method and were studied their ability for removing hexadecane in liquid and solid medium. According to results, the isolated bacteria were identified as O.oryzae and P.lautus. In liquid medium, hexadecane concentration decreased from 3000±1.4 to 366.96±0.9 mg/l (87.77±0.2) by O.oryzae, while the removal percent by P.lautus was 80.89±1.2 after 33 days. Hexadecane concentration decreased from 30 to 18.09±0.6 g/kg soil (39.69±1.1) during 80 days. The pseudo-first order was the best model for biodegradation by both cultures. Whereas, the kinetic behavior of hexadecane bioremediation was described by first and pseudo-first-order model (r2 = 0.798). In addition, these isolated bacteria have higher efficiency in hexadecane removal in comparison to the other previously identified bacteria. Moreover, O.oryzae can be used in remediation of petroleum products, especially diesel oil. © 2020, © 2020 Taylor & Francis

In vitro cytotoxic and apoptotic activities of Allium paradoxum (M. bieb.) G. Don extract on human breast cancer cell line

Researchers from all pharmaceutical fields are trying to find new drugs from natural origin with less toxicity. In northern Hyrcanian forests Iran, Allium paradoxum (M. Bieb.) G. Don has traditionally used as food and vegetable. Previously studies reports, this plant has a medicinal potential for anti-oxidant and anti-hemolytic activities. In this regard, we evaluated the anti-tumor activity of hydroalcoholic extract of A. paradoxum (M. Bieb.) G. Don in different concentrations on human breast cancer cell line (MCF-7). MTT assay was performed with MCF-7 cancer cell line and also evaluation of apoptotic effect, Bax and Bcl-2 expression in MCF7 cells were analyzed by real time RT-PCR. The results showed that the A. paradoxum (M. Bieb.) G. Don extracts decrease the viability of MCF-7 cell line in a dose-dependent manner and the most effective concentration of this extracts after 24 h treatment was 100 µM. Apoptosis induction was confirmed by fluorescence microscopy and plant extracts display a pro-apoptotic effect by down-regulated and up-regulated the expression of Bcl-2 and BAX in tumor cells, respectively. In conclusion, the study was confirmed pro-apoptotic and cytotoxicity effect of A. paradoxum (M. Bieb.) G. Don extract against MCF-7 cell lines. Based on being natural, low cost, accessibility, and noteworthy advantages of this product, it seems that A. paradoxum (M. Bieb.) G. Don has a potential source for isolation of novel anticancer agents for a drug. © 2018, National Institute of Science Communication and Information Resources (NISCAIR). All rights reserved

Urinary levels of potentially toxic elements (PTEs) in female beauticians and their association with urinary biomarkers of oxidative stress/inflammation and kidney injury

The present research was aimed to assess the urinary levels of potentially toxic elements (PTEs) in female beauticians and its correlation with oxidative stress/inflammation and kidney injury. To this end, the urine samples were collected from 50 female beauticians from beauty salons (exposed group) and 35 housewives (control group), and then, the level of PTEs was determined. The mean levels of the sum of urinary PTEs (∑PTEs) biomarkers in before and after exposure and control group were 83.55, 114.27 and 13.61 μg/L, respectively. Results also showed that the urinary level of PTEs biomarkers is significantly higher in women occupationally exposed to cosmetics compared to control group. The urinary levels of arsenic (As), cadmium (Cd), lead (Pb), and chromium (Cr) biomarkers have high correlation coefficients with early oxidative stress effects such as 8-Hydroxyguanosine (8-OHdG), 8-isoprostane and Malondialdehyde (MDA). Moreover, As and Cd biomarker levels were positively and significantly associated with kidney damages such as urinary kidney injury molecule-1 (uKIM-1) and tissue inhibitor matrix metalloproteinase 1 (uTIMP-1) (P < 0.01). Therefore, women who working in beauty salons can probably be categorized as high – exposure and high-risk workers in terms of DNA oxidative and kidney damages

Global, regional, and national sex differences in the global burden of tuberculosis by HIV status, 1990–2019: results from the Global Burden of Disease Study 2019

Background: Tuberculosis is a major contributor to the global burden of disease, causing more than a million deaths annually. Given an emphasis on equity in access to diagnosis and treatment of tuberculosis in global health targets, evaluations of differences in tuberculosis burden by sex are crucial. We aimed to assess the levels and trends of the global burden of tuberculosis, with an emphasis on investigating differences in sex by HIV status for 204 countries and territories from 1990 to 2019. Methods: We used a Bayesian hierarchical Cause of Death Ensemble model (CODEm) platform to analyse 21 505 site-years of vital registration data, 705 site-years of verbal autopsy data, 825 site-years of sample-based vital registration data, and 680 site-years of mortality surveillance data to estimate mortality due to tuberculosis among HIV-negative individuals. We used a population attributable fraction approach to estimate mortality related to HIV and tuberculosis coinfection. A compartmental meta-regression tool (DisMod-MR 2.1) was then used to synthesise all available data sources, including prevalence surveys, annual case notifications, population-based tuberculin surveys, and tuberculosis cause-specific mortality, to produce estimates of incidence, prevalence, and mortality that were internally consistent. We further estimated the fraction of tuberculosis mortality that is attributable to independent effects of risk factors, including smoking, alcohol use, and diabetes, for HIV-negative individuals. For individuals with HIV and tuberculosis coinfection, we assessed mortality attributable to HIV risk factors including unsafe sex, intimate partner violence (only estimated among females), and injection drug use. We present 95% uncertainty intervals for all estimates. Findings: Globally, in 2019, among HIV-negative individuals, there were 1·18 million (95% uncertainty interval 1·08–1·29) deaths due to tuberculosis and 8·50 million (7·45–9·73) incident cases of tuberculosis. Among HIV-positive individuals, there were 217 000 (153 000–279 000) deaths due to tuberculosis and 1·15 million (1·01–1·32) incident cases in 2019. More deaths and incident cases occurred in males than in females among HIV-negative individuals globally in 2019, with 342 000 (234 000–425 000) more deaths and 1·01 million (0·82–1·23) more incident cases in males than in females. Among HIV-positive individuals, 6250 (1820–11 400) more deaths and 81 100 (63 300–100 000) more incident cases occurred among females than among males in 2019. Age-standardised mortality rates among HIV-negative males were more than two times greater in 105 countries and age-standardised incidence rates were more than 1·5 times greater in 74 countries than among HIV-negative females in 2019. The fraction of global tuberculosis deaths among HIV-negative individuals attributable to alcohol use, smoking, and diabetes was 4·27 (3·69–5·02), 6·17 (5·48–7·02), and 1·17 (1·07–1·28) times higher, respectively, among males than among females in 2019. Among individuals with HIV and tuberculosis coinfection, the fraction of mortality attributable to injection drug use was 2·23 (2·03–2·44) times greater among males than females, whereas the fraction due to unsafe sex was 1·06 (1·05–1·08) times greater among females than males. Interpretation: As countries refine national tuberculosis programmes and strategies to end the tuberculosis epidemic, the excess burden experienced by males is important. Interventions are needed to actively communicate, especially to men, the importance of early diagnosis and treatment. These interventions should occur in parallel with efforts to minimise excess HIV burden among women in the highest HIV burden countries that are contributing to excess HIV and tuberculosis coinfection burden for females. Placing a focus on tuberculosis burden among HIV-negative males and HIV and tuberculosis coinfection among females might help to diminish the overall burden of tuberculosis. This strategy will be crucial in reaching both equity and burden targets outlined by global health milestones. Funding: Bill & Melinda Gates Foundation. © 2022 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 licens

Spatial, temporal, and demographic patterns in prevalence of smoking tobacco use and attributable disease burden in 204 countries and territories, 1990�2019: a systematic analysis from the Global Burden of Disease Study 2019

Background: Ending the global tobacco epidemic is a defining challenge in global health. Timely and comprehensive estimates of the prevalence of smoking tobacco use and attributable disease burden are needed to guide tobacco control efforts nationally and globally. Methods: We estimated the prevalence of smoking tobacco use and attributable disease burden for 204 countries and territories, by age and sex, from 1990 to 2019 as part of the Global Burden of Diseases, Injuries, and Risk Factors Study. We modelled multiple smoking-related indicators from 3625 nationally representative surveys. We completed systematic reviews and did Bayesian meta-regressions for 36 causally linked health outcomes to estimate non-linear dose-response risk curves for current and former smokers. We used a direct estimation approach to estimate attributable burden, providing more comprehensive estimates of the health effects of smoking than previously available. Findings: Globally in 2019, 1·14 billion (95 uncertainty interval 1·13�1·16) individuals were current smokers, who consumed 7·41 trillion (7·11�7·74) cigarette-equivalents of tobacco in 2019. Although prevalence of smoking had decreased significantly since 1990 among both males (27·5 26·5�28·5 reduction) and females (37·7% 35·4�39·9 reduction) aged 15 years and older, population growth has led to a significant increase in the total number of smokers from 0·99 billion (0·98�1·00) in 1990. Globally in 2019, smoking tobacco use accounted for 7·69 million (7·16�8·20) deaths and 200 million (185�214) disability-adjusted life-years, and was the leading risk factor for death among males (20·2% 19·3�21·1 of male deaths). 6·68 million 86·9% of 7·69 million deaths attributable to smoking tobacco use were among current smokers. Interpretation: In the absence of intervention, the annual toll of 7·69 million deaths and 200 million disability-adjusted life-years attributable to smoking will increase over the coming decades. Substantial progress in reducing the prevalence of smoking tobacco use has been observed in countries from all regions and at all stages of development, but a large implementation gap remains for tobacco control. Countries have a clear and urgent opportunity to pass strong, evidence-based policies to accelerate reductions in the prevalence of smoking and reap massive health benefits for their citizens. Funding: Bloomberg Philanthropies and the Bill & Melinda Gates Foundation. © 2021 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 licens