Construction of pLLO vector encoding truncated form of Listeriolysin O as molecular adjuvant for DNA vaccine studies

Background: The major problem of DNA vaccine is less immunogenicity of them verses other killed or live whole organism vaccines therefore adjuvants for use in this kind vaccines is very necessary. Genetic adjuvants with bacterial sources are an appropriate approach to modulate immune responses to DNA vaccines. Listeria Monocytogenes proteins such as Listeriolysin O (LLO) with CD4 and CD8 epitopes can be as an adjuvant to initiate both innate and adaptive immune responses if the protein cytotoxicity can be eliminated. Herein we constructed a truncated LLO plasmid as genetic adjuvant and tested it in combination with a DNA construct as a model vaccine.Materials and Methods: About 1340bp of the 5' end of whole LLO gene was amplified by PCR on DNA purified from Listeria Monocytogenes. Sequential sub cloning of truncated LLO into the Xho I/EcoRV sites of pcDNA3.1 plasmid, downstream of CMV promoter was done. pLLO plasmid was transfected to HEK293T cell line by lipofection method. LLO protein expression from transiently transfected 293T cell lysates was confirmed by western blotting. Then the adjuvant activity of LLO in BALB/c mice model was analyzed using proliferation test.Results: Double digestion of pLLO plasmid with the enzymes that were applied for cloning led to the isolation of two fragments with expected sizes. The final plasmid was also confirmed following sequencing reactions. Moreover, expression of LLO was evidenced in transfected 293T cells, compared to non-transfected controls. In vivo study was shown, high significant proliferative responses in LLO co-immunization pattern.Conclusion: In the DNA vaccine study, LLO co-administration plasmid could be a suitable genetic adjuvant to enhance cellular immune response of vaccine

A potential hypothesis for 2019-nCoV infection therapy through delivery of recombinant ACE2 by red blood cell-hitchhiking

A novel infectious disease, caused by 2019 Novel Coronavirus (2019-nCoV) is responsible for the recent outbreak of severe respiratory disease. The 2019-nCoV spread rapidly and reaching epidemic proportions in many countries of the world. ACE2 was identified as a key receptor for 2019-nCoV infections. Excessive form of soluble ACE2 rescues cellular ACE2 activity which has a protective role in acute lung failure and neutralizes the virus. The short half-life of ACE2 is a major limitation to its practical application. Nanoparticle-based drug delivery systems are one of the most widely investigated approaches for developing novel therapies for a variety of diseases. Nevertheless, nanoparticles suffer from the rapid removal from the bloodstream by the reticuloendothelial system (RES). A noncovalent attachment of nanoparticles to RBCs increases their half-life in blood and allows transient accumulation in the lungs, while decreases their uptake by the liver and spleen. Connecting the recombinant ACE2 into the surface of nanoparticles that were attached to RBCs can be a potential therapeutic approach for 2019-nCoV infection through increasing their lung targeting to naturalize the virus and also acting as a bioreactor in the blood circulation to decrease serum level of Angiotensin II and protects lungs from injury/ARDS

New Rule Base Business Success Prediction Model for Iranian IT Start-Ups

Hundreds of thousands of new businesses are created every year around the world, and it is estimated that about half a billion people worldwide are actively trying to start new businesses. Therefore, creating a new business is very important; because it creates new job opportunities, produces new technology and creates wealth and value in society. However, a large percentage of small businesses fail in the early years of their existence. Meanwhile, in the field of superior technologies, due to the high dependence on changing technology and also the need for high initial capital, firms are facing a more difficult situation. The purpose of this study is to provide a rule based database to determine the success and failure of IT startups in Iran. For this purpose, using research conducted in this field and also receiving the opinions of experts through a semi-open questionnaire, we identified 36 factors affecting the success and failure of startups and classified them into 5 categories. In this research, the method of fuzzy inference system by Mamdani method has been used to analyze the factors. The statistical community is the founders of Iranian startups as well as professors and business professionals;30 founders of successful Iranian startups have been randomly selected as a sample. The results show that team-related factors and team characteristics have the greatest impact on the success and failure of startups

Gd 3+

Designing a unique theranostic biocompatible, biodegradable, and cost-effective agent which is easy to be synthesized as a biohybrid material was the aim of this study. In this matter, asparagine attached to anionic linear globular dendrimer G2 (as a biocompatible, biodegradable, and cost-effective agent which is negatively charged nanosized and water soluble polymer that outweighs other traditionally used dendrimers) and finally contrast agent (Gd3+) was loaded (which made complexes) in synthesized asparagine-dendrimer. Observations revealed that, in addition to successful colon cancer and brain targeting, Gd3+-dendrimer-asparagine, the proposed theranostic agent, could increase T1 MR relaxation times, decrease T2 MR relaxation times significantly, and improve contrast of image as well as illustrating good cellular uptake based on florescent microscopy/flow cytometry and ICP-mass data. In addition to that, it increased tumor growth inhibition percentage (TGI%) significantly compared to FDA approved contrast agent, Magnevist. Totally, Gd3+-anionic linear globular dendrimer G2-asparagine could be introduced to the cancer imaging/therapy (theranostics) protocols after in vivo MR and fluorescent analysis and passing clinical trials. Hence, this nanotheranostic agent would be a promising candidate for brain drug delivery and imaging in the future

Improvement of Mouse Preantral Follicle Survival and Development following Co-Culture with Ovarian Parenchyma Cell Suspension

Background: The parallel and continued improvements in both infertility treatment and the management of malignancycases have brought to the forefront the potential for fertility preservation. Using ovarian follicular resourcescan effectively improve reproductive capacity and prevent infertility. The primary aim of this research was to try togenerate an appropriate in vivo environment for the growth of the mouse follicles. Hence, the possible effects of theovarian parenchyma cell suspension were explored on the growth and maturation of preantral follicles in vitro.Materials and Methods: In this experimental study, ovarian parenchymal cells were mechanically dissociated frompreantral follicles of 12-14 days-old NMRI mice and then divided into 5 experimental groups (G1: Control, G2: Freshfollicle with fresh parenchyma cell suspension, G3: Vitrified-warmed follicle with fresh parenchyma cell suspension,G4: Fresh follicle with frozen-thawed parenchyma cell suspension, and G5: Vitrified-warmed follicle with frozen-thawedparenchyma cell suspension). The diameter of the follicles and immature oocytes, viability, antrum formation,resumption of meiosis, in vitro fertilization (IVF), and Gdf9, Bmp6, and Bmp15 gene expression were examined ondifferent periods.Results: The diameter of the follicles and the oocytes on days 4 and 8, as well as the survival rate of the follicles upto day 12, were significantly higher in G2 and G4 compared to the Ctrl group (G1: 73.66%, G2:87.99%, G3: 82.70%,G4: 94.37%, and G5: 78.59%). Expression of growth marker genes for G3, and G5 groups was significantly higherthan other groups, which indicated the protective effects of parenchyma cell suspension on follicles damaged by vitrificationsolutions.Conclusion: The growth, survival, and maturation of preantral follicles could be enhanced by co-culturing them withovarian parenchyma cells. Further studies are needed to optimize the conditions for a successful parenchyma cellsuspension-induced in vitro maturation (IVM) to occur in infertility clinics

Pulmonary manifestations in a cohort of patients with inborn errors of immunity : an 8-year follow-up study

Background: Inborn errors of immunity (IEIs) are a group of congenital diseases caused by genetic defects in the development and function of the immune system. The involvement of the respiratory tract is one of the most common presentations in IEIs. Methods: Overall, 117 patients with diagnosed IEIs were followed-up within 8 years at the National Research Institute of Tuberculosis and Lung Diseases (NRITLD). Demographic, clinical, and laboratory data were collected in a questionnaire. Pulmonary function test (PFT), chest X-ray (CXR), and high-resolution computed tomography (HRCT) scans were obtained where applicable. Results: Our study population consisted of 48 (41%) patients with predominantly antibody deficiencies (PADs), 39 (32%) patients with congenital defects of phagocytes, 14 (11.9%) patients with combined immunodeficiency (CID), and 16 (14%) patients with Mendelian susceptibility to mycobacterial diseases (MSMD).. Recurrent pneumonia was the most common manifestation, while productive cough appeared to be the most common symptom in almost all diseases. PFT showed an obstructive pattern in patients with PAD, a restrictive pattern in patients with CID, and a mixed pattern in patients with CGD. HRCT findings were consistent with bronchiectasis in most PAD patients, whereas consolidation and mediastinal lesions were more common in the other groups. Conclusions: Pulmonary manifestations vary among different groups of IEIs. The screening for lung complications should be performed regularly to reveal respiratory pathologies in early stages and follow-up on already existing abnormalities. (C) 2022 Codon Publications. Published by Codon Publications.Peer reviewe

Prevalence and Correlates of Psychiatric Disorders in a National Survey of Iranian Children and Adolescents

Objective: Considering the impact of rapid sociocultural, political, and economical changes on societies and families, population-based surveys of mental disorders in different communities are needed to describe the magnitude of mental health problems and their disabling effects at the individual, familial, and societal levels. Method: A population-based cross sectional survey (IRCAP project) of 30 532 children and adolescents between 6 and 18 years was conducted in all provinces of Iran using a multistage cluster sampling method. Data were collected by 250 clinical psychologists trained to use the validated Persian version of the semi-structured diagnostic interview Kiddie-Schedule for Affective Disorders and Schizophrenia-PL (K-SADS-PL). Results: In this national epidemiological survey, 6209 out of 30 532 (22.31%) were diagnosed with at least one psychiatric disorder. The anxiety disorders (14.13%) and behavioral disorders (8.3%) had the highest prevalence, while eating disorders (0.13%) and psychotic symptoms (0.26%) had the lowest. The prevalence of psychiatric disorders was significantly lower in girls (OR = 0.85; 95% CI: 0.80-0.90), in those living in the rural area (OR = 0.80; 95% CI: 0.73-0.87), in those aged 15-18 years (OR = 0.92; 95% CI: 0.86-0.99), as well as that was significantly higher in those who had a parent suffering from mental disorders (OR = 1.96; 95% CI: 1.63-2.36 for mother and OR = 1.33; 95% CI: 1.07-1.66 for father) or physical illness (OR = 1.26; 95% CI: 1.17-1.35 for mother and OR = 1.19; 95% CI: 1.10-1.28 for father). Conclusion: About one fifth of Iranian children and adolescents suffer from at least one psychiatric disorder. Therefore, we should give a greater priority to promoting mental health and public health, provide more accessible services and trainings, and reduce barriers to accessing existing services

Future and potential spending on health 2015-40 : development assistance for health, and government, prepaid private, and out-of-pocket health spending in 184 countries

Background The amount of resources, particularly prepaid resources, available for health can affect access to health care and health outcomes. Although health spending tends to increase with economic development, tremendous variation exists among health financing systems. Estimates of future spending can be beneficial for policy makers and planners, and can identify financing gaps. In this study, we estimate future gross domestic product (GDP), all-sector government spending, and health spending disaggregated by source, and we compare expected future spending to potential future spending. Methods We extracted GDP, government spending in 184 countries from 1980-2015, and health spend data from 1995-2014. We used a series of ensemble models to estimate future GDP, all-sector government spending, development assistance for health, and government, out-of-pocket, and prepaid private health spending through 2040. We used frontier analyses to identify patterns exhibited by the countries that dedicate the most funding to health, and used these frontiers to estimate potential health spending for each low-income or middle-income country. All estimates are inflation and purchasing power adjusted. Findings We estimated that global spending on health will increase from US$9.21 trillion in 2014 to$24.24 trillion (uncertainty interval [UI] 20.47-29.72) in 2040. We expect per capita health spending to increase fastest in upper-middle-income countries, at 5.3% (UI 4.1-6.8) per year. This growth is driven by continued growth in GDP, government spending, and government health spending. Lower-middle income countries are expected to grow at 4.2% (3.8-4.9). High-income countries are expected to grow at 2.1% (UI 1.8-2.4) and low-income countries are expected to grow at 1.8% (1.0-2.8). Despite this growth, health spending per capita in low-income countries is expected to remain low, at $154 (UI 133-181) per capita in 2030 and$195 (157-258) per capita in 2040. Increases in national health spending to reach the level of the countries who spend the most on health, relative to their level of economic development, would mean $321 (157-258) per capita was available for health in 2040 in low-income countries. Interpretation Health spending is associated with economic development but past trends and relationships suggest that spending will remain variable, and low in some low-resource settings. Policy change could lead to increased health spending, although for the poorest countries external support might remain essential.Peer reviewe

Mapping 123 million neonatal, infant and child deaths between 2000 and 2017

Since 2000, many countries have achieved considerable success in improving child survival, but localized progress remains unclear. To inform efforts towards United Nations Sustainable Development Goal 3.2—to end preventable child deaths by 2030—we need consistently estimated data at the subnational level regarding child mortality rates and trends. Here we quantified, for the period 2000–2017, the subnational variation in mortality rates and number of deaths of neonates, infants and children under 5 years of age within 99 low- and middle-income countries using a geostatistical survival model. We estimated that 32% of children under 5 in these countries lived in districts that had attained rates of 25 or fewer child deaths per 1,000 live births by 2017, and that 58% of child deaths between 2000 and 2017 in these countries could have been averted in the absence of geographical inequality. This study enables the identification of high-mortality clusters, patterns of progress and geographical inequalities to inform appropriate investments and implementations that will help to improve the health of all populations