Recombinant production of Soluble Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (sTRAIL) as a therapeutic protein

Abstract:  Successfully cancer therapies aim to induce apoptosis in cancer cell lines. Recent advances in cancer therapy based on the use of some recombinant proteins such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). TRAIL is a new member of the TNF superfamily. In this paper, we report the expression, purification, and  preparation of a recombinant form of the extracelluar domain of the TRAIL (sTRAIL) in Escherichia coli rosetta gami under the control of T7 promoter; which  may selectively induce apoptosis of tumor cells in vitro. To obtain recombinant sTRAIL protein, the encoding region for sTRAIL was cloned between Xho1 and BamHI in pET28a expression vector. The results showed that the recombinant sTRAIL was efficiently produced in Ecoli rosetta gami strain.                               Introduction: Apoptosis is an evolutionarily conserved and essential for maintenance of  tissue homeostasis and removal of unwanted cells. TRAIL belongs to the group of therapeutic agents selectively targeting a wide variety of cancer cells without affecting the normal cells .The therapeutic potential of TRAIL is attributed to its receptor expression in a variety of tissues; which initiates apoptosis in cancer cells through interaction with the death receptors DR4 and DR5. Due to its selective nature, it is considered as a significant therapeutic agent in cancer therapy. The purpose of this study was to produce recombinant human sTRAIL in Rosetta Gami2 E.coli strain and its functions on cancerous cells in vitro. Methods and results:  we optimized the coding sequence of this protein. The recombinant plasmid was transformed into Rosetta Gami2 E.coli strain for expression. The transformed bacteria which contain recombinant plasmid were cultured in 37ºC with 250 rpm in LB and in 20ºC in TB medium for 18 hours. TRAIL was purified by Ni sepharose column, and the presence of the recombinant protein was confirmed by SDS-PAGE. The concentration of purified protein was measured by Bradford assay. Our finding showed that the recombinant protein (34kD) has been successfully produced for next experiments, the purified protein was desalted and applied toward cancerous cells. Conclusions: In summary, TRAIL can be considered as a promising therapeutic agent for effective, targeted and less toxic agents for treatment of cancers

KLINIČKE PRIČE I MEDICINSKE ANAMNEZE KOJE JE ZABILJEŽIO RHAZES (865.–925.), IRANSKO‑ISLAMSKI LIJEČNIK SREDNJEGA VIJEKA

Recording medical histories of patients is not a new issue in clinical medicine. However, the method practiced by the Iranian chemist physician, Rhazes, in the ninth century A.D is incredible. Rhazes has written several textbooks in clinical medicine, but a particular one, “Clinical Stories and Medical Histories” (Qesas va hekayat al-marazi), is a classical case book describing precise clinical courses of thirty three patients. Each chapter includes a title, the name and demographic data about a patient, his/her history of present illness, past medical and family history, findings of physical exam, impression and interventions by the physician, including pharmacological or surgical management. The reasons for each decision made by Rhazes as well as the outcomes are clearly discussed. This book review will shed light on the unknown medical practice methods in Islamic-Iranian golden era.Bilježenje medicinske anamneze pacijenata nije novo pitanje kliničke medicine. Kako bilo, metoda koju je prakticirao Rhazes, iranski kemičar i liječnik, u devetom stoljeću zadivljujuća je. Rhazes je napisao nekoliko udžbenika iz kliničke medicine, ali ona kratka, Kliničke priče i medicinske anamneze, klasična je knjiga prikaza slučajeva koja opisuje točan klinički tijek kod trideset i tri pacijenta. Svako poglavlje uključuje naslov, ime i demografske podatke o pacijentu, njenu/njegovu povijest trenutne bolesti, prošlu medicinsku i obiteljsku povijest bolesti, nalaze fizikalnog pregleda, utiske i intervencije liječnika, uključujući farmakološke i/ili kirurške postupke. Razlozi su svake Rhazesove odluke kao i rezultati jasno raspravljeni. Čitanje zabilješki u ovoj knjizi pomaže osvijetliti nepoznate kutke koji se tiču metoda medicinskih praksi u vrijeme rađanja islamskoiranske civilizacije

Associations between Potential Inflammatory Properties of the Diet and Frequency, Duration, and Severity of Migraine Headaches: A Cross-Sectional Study

Despite a large body of literature on the association between the dietary inflammatory index (DII) and various chronic diseases, limited knowledge is available regarding the association between DII and migraine. Therefore, we assessed the relationship between the DII and migraine characteristics, including duration, frequency, and severity of migraine headaches, Headache Impact Test-6 (HIT-6), and serum levels of nitric oxide (NO). This population-based cross-sectional study was conducted from August 2019 to June 2020 among 262 patients (38 men and 224 women; 20-50 years). A 168-item semiquantitative food frequency questionnaire (FFQ) was gathered to evaluate dietary intake, and subsequently, an energy-adjusted DII score was calculated. After controlling for potential confounders, an increase of 3.48 in headache frequency was observed when the DII score increased from - 4.04 to - 1.83 (β = 3.48; 95% CI 1.43, 5.54). In the crude model, headache duration tended to be inversely associated with DII in the subjects with the pro-inflammatory diet compared to those with the anti-inflammatory diet (β = - 0.22; 95% CI - 0.46, 0.02). After adjustment for confounders, those with the highest DII values were at a higher risk of severe headaches than those with the lowest values (OR = 2.25; 95% CI 1.17, 4.32). No other significant results were found in terms of the association between DII and HIT-6 or serum NO levels. We found evidence suggesting that higher adherence to a diet with anti-inflammatory properties was significantly and inversely related to headache frequency. Furthermore, our results suggest that the DII score is substantially related to migraine severity

Comparing the Effects of Aquatic and Land-based Exercises on the Bio-motor Abilities of Elderly Men

Background: Physical activity, whether in water or on land, can improve motor disorders in the elderly. However, the more effective type of exercise remains inconspicuous. Aim: To compare the effects of aquatic and land-based exercises on the bio-motor abilities of elderly men. Method: This clinical trial was performed on 60 elderly men visiting Imamat and Daneshamuz health centers in Mashhad, Iran, in 2016. The eligible volunteers were randomly assigned to two groups (n=30 each). In one group, the subjects attended aquatic exercise sessions for six weeks, while the other group participated a land-based exercise program. Bio-motor abilities of the subjects including motor ability, flexibility, and general endurance were evaluated before and immediately after the intervention and recorded in a researcher-made checklist. Data were analyzed in SPSS, version 16, using the independent t-test, paired t-test, Chi-square test, Fisher’s exact test, and exact Chi-square test. Results: The mean ages of the aquatic exercise and land-based exercise groups were respectively 69.9±5.1 and 68.0±5.0 years. Post-intervention, the aquatic exercise group showed significantly higher motor ability (P=0.01) and general endurance (P=0.02) than did the other group. However, the difference in final flexibility of the groups was not significant (P=0.252). Paired t-test showed a significant improvement in bio-motor abilities of both groups post-intervention (

Comparison ability of algae and nanoparticles on nitrate and phosphate removal from aquaculture wastewater

Background: Aquaculture wastewater contains high levels of phosphate and nitrate. The reuse of this water requires standards beyond the secondary standards to eliminate more organic pollutants from aquaculture effluents. In this research, the removal of these pollutants from wastewater using Chlorella vulgaris and Fe3O4 nanoparticles in the reactor space was investigated. Methods: This study was conducted on fish farms effluent in the laboratory system. For this purpose, a 5-L semi-industrial reactor with a mixer blade, porous plate, and a compressor was designed. Chlorella vulgaris samples were collected from the natural environment and cultured in the laboratory environment. Also, Fe3O4 nanoparticles were prepared from Iranian Nano Pishgaman Company to make the desired solution. During the experiment (3 weeks), samples were taken weekly (in one phase) from the effluent. Dissolved oxygen (DO), pH, nitrate (NO3), and phosphate (PO4) factors from the influent and effluent of the farms were measured. The statistical data were analyzed using SPSS version 21 and Excel 2013. Results: The amounts of nitrate and phosphate were decreased by about 80.76 and 80.55% in the biological reactor, whereas these amounts were 70.52 and 70.48% in the nanoparticle reactor, respectively. Also, there were significant differences in the amounts of NO3 and PO4 between the control treatment and weekly treatment (P < 0.05). Conclusion: Based on the results, both reactors were able to reduce nitrate and phosphate from aquaculture wastewater, but the efficiency of the biological reactor was higher than that of the nanoparticle reactor. Keywords: Nitrate, Phosphate, Fe3O4 nanoparticle, Chlorella vulgaris, Biological refinemen

Global age-sex-specific mortality, life expectancy, and population estimates in 204 countries and territories and 811 subnational locations, 1950–2021, and the impact of the COVID-19 pandemic: a comprehensive demographic analysis for the Global Burden of Disease Study 2021

Background: Estimates of demographic metrics are crucial to assess levels and trends of population health outcomes. The profound impact of the COVID-19 pandemic on populations worldwide has underscored the need for timely estimates to understand this unprecedented event within the context of long-term population health trends. The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 provides new demographic estimates for 204 countries and territories and 811 additional subnational locations from 1950 to 2021, with a particular emphasis on changes in mortality and life expectancy that occurred during the 2020–21 COVID-19 pandemic period. Methods: 22 223 data sources from vital registration, sample registration, surveys, censuses, and other sources were used to estimate mortality, with a subset of these sources used exclusively to estimate excess mortality due to the COVID-19 pandemic. 2026 data sources were used for population estimation. Additional sources were used to estimate migration; the effects of the HIV epidemic; and demographic discontinuities due to conflicts, famines, natural disasters, and pandemics, which are used as inputs for estimating mortality and population. Spatiotemporal Gaussian process regression (ST-GPR) was used to generate under-5 mortality rates, which synthesised 30 763 location-years of vital registration and sample registration data, 1365 surveys and censuses, and 80 other sources. ST-GPR was also used to estimate adult mortality (between ages 15 and 59 years) based on information from 31 642 location-years of vital registration and sample registration data, 355 surveys and censuses, and 24 other sources. Estimates of child and adult mortality rates were then used to generate life tables with a relational model life table system. For countries with large HIV epidemics, life tables were adjusted using independent estimates of HIV-specific mortality generated via an epidemiological analysis of HIV prevalence surveys, antenatal clinic serosurveillance, and other data sources. Excess mortality due to the COVID-19 pandemic in 2020 and 2021 was determined by subtracting observed all-cause mortality (adjusted for late registration and mortality anomalies) from the mortality expected in the absence of the pandemic. Expected mortality was calculated based on historical trends using an ensemble of models. In location-years where all-cause mortality data were unavailable, we estimated excess mortality rates using a regression model with covariates pertaining to the pandemic. Population size was computed using a Bayesian hierarchical cohort component model. Life expectancy was calculated using age-specific mortality rates and standard demographic methods. Uncertainty intervals (UIs) were calculated for every metric using the 25th and 975th ordered values from a 1000-draw posterior distribution. Findings: Global all-cause mortality followed two distinct patterns over the study period: age-standardised mortality rates declined between 1950 and 2019 (a 62·8% [95% UI 60·5–65·1] decline), and increased during the COVID-19 pandemic period (2020–21; 5·1% [0·9–9·6] increase). In contrast with the overall reverse in mortality trends during the pandemic period, child mortality continued to decline, with 4·66 million (3·98–5·50) global deaths in children younger than 5 years in 2021 compared with 5·21 million (4·50–6·01) in 2019. An estimated 131 million (126–137) people died globally from all causes in 2020 and 2021 combined, of which 15·9 million (14·7–17·2) were due to the COVID-19 pandemic (measured by excess mortality, which includes deaths directly due to SARS-CoV-2 infection and those indirectly due to other social, economic, or behavioural changes associated with the pandemic). Excess mortality rates exceeded 150 deaths per 100 000 population during at least one year of the pandemic in 80 countries and territories, whereas 20 nations had a negative excess mortality rate in 2020 or 2021, indicating that all-cause mortality in these countries was lower during the pandemic than expected based on historical trends. Between 1950 and 2021, global life expectancy at birth increased by 22·7 years (20·8–24·8), from 49·0 years (46·7–51·3) to 71·7 years (70·9–72·5). Global life expectancy at birth declined by 1·6 years (1·0–2·2) between 2019 and 2021, reversing historical trends. An increase in life expectancy was only observed in 32 (15·7%) of 204 countries and territories between 2019 and 2021. The global population reached 7·89 billion (7·67–8·13) people in 2021, by which time 56 of 204 countries and territories had peaked and subsequently populations have declined. The largest proportion of population growth between 2020 and 2021 was in sub-Saharan Africa (39·5% [28·4–52·7]) and south Asia (26·3% [9·0–44·7]). From 2000 to 2021, the ratio of the population aged 65 years and older to the population aged younger than 15 years increased in 188 (92·2%) of 204 nations. Interpretation: Global adult mortality rates markedly increased during the COVID-19 pandemic in 2020 and 2021, reversing past decreasing trends, while child mortality rates continued to decline, albeit more slowly than in earlier years. Although COVID-19 had a substantial impact on many demographic indicators during the first 2 years of the pandemic, overall global health progress over the 72 years evaluated has been profound, with considerable improvements in mortality and life expectancy. Additionally, we observed a deceleration of global population growth since 2017, despite steady or increasing growth in lower-income countries, combined with a continued global shift of population age structures towards older ages. These demographic changes will likely present future challenges to health systems, economies, and societies. The comprehensive demographic estimates reported here will enable researchers, policy makers, health practitioners, and other key stakeholders to better understand and address the profound changes that have occurred in the global health landscape following the first 2 years of the COVID-19 pandemic, and longer-term trends beyond the pandemic

Global incidence, prevalence, years lived with disability (YLDs), disability-adjusted life-years (DALYs), and healthy life expectancy (HALE) for 371 diseases and injuries in 204 countries and territories and 811 subnational locations, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

Background: Detailed, comprehensive, and timely reporting on population health by underlying causes of disability and premature death is crucial to understanding and responding to complex patterns of disease and injury burden over time and across age groups, sexes, and locations. The availability of disease burden estimates can promote evidence-based interventions that enable public health researchers, policy makers, and other professionals to implement strategies that can mitigate diseases. It can also facilitate more rigorous monitoring of progress towards national and international health targets, such as the Sustainable Development Goals. For three decades, the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) has filled that need. A global network of collaborators contributed to the production of GBD 2021 by providing, reviewing, and analysing all available data. GBD estimates are updated routinely with additional data and refined analytical methods. GBD 2021 presents, for the first time, estimates of health loss due to the COVID-19 pandemic. Methods: The GBD 2021 disease and injury burden analysis estimated years lived with disability (YLDs), years of life lost (YLLs), disability-adjusted life-years (DALYs), and healthy life expectancy (HALE) for 371 diseases and injuries using 100 983 data sources. Data were extracted from vital registration systems, verbal autopsies, censuses, household surveys, disease-specific registries, health service contact data, and other sources. YLDs were calculated by multiplying cause-age-sex-location-year-specific prevalence of sequelae by their respective disability weights, for each disease and injury. YLLs were calculated by multiplying cause-age-sex-location-year-specific deaths by the standard life expectancy at the age that death occurred. DALYs were calculated by summing YLDs and YLLs. HALE estimates were produced using YLDs per capita and age-specific mortality rates by location, age, sex, year, and cause. 95% uncertainty intervals (UIs) were generated for all final estimates as the 2·5th and 97·5th percentiles values of 500 draws. Uncertainty was propagated at each step of the estimation process. Counts and age-standardised rates were calculated globally, for seven super-regions, 21 regions, 204 countries and territories (including 21 countries with subnational locations), and 811 subnational locations, from 1990 to 2021. Here we report data for 2010 to 2021 to highlight trends in disease burden over the past decade and through the first 2 years of the COVID-19 pandemic. Findings: Global DALYs increased from 2·63 billion (95% UI 2·44–2·85) in 2010 to 2·88 billion (2·64–3·15) in 2021 for all causes combined. Much of this increase in the number of DALYs was due to population growth and ageing, as indicated by a decrease in global age-standardised all-cause DALY rates of 14·2% (95% UI 10·7–17·3) between 2010 and 2019. Notably, however, this decrease in rates reversed during the first 2 years of the COVID-19 pandemic, with increases in global age-standardised all-cause DALY rates since 2019 of 4·1% (1·8–6·3) in 2020 and 7·2% (4·7–10·0) in 2021. In 2021, COVID-19 was the leading cause of DALYs globally (212·0 million [198·0–234·5] DALYs), followed by ischaemic heart disease (188·3 million [176·7–198·3]), neonatal disorders (186·3 million [162·3–214·9]), and stroke (160·4 million [148·0–171·7]). However, notable health gains were seen among other leading communicable, maternal, neonatal, and nutritional (CMNN) diseases. Globally between 2010 and 2021, the age-standardised DALY rates for HIV/AIDS decreased by 47·8% (43·3–51·7) and for diarrhoeal diseases decreased by 47·0% (39·9–52·9). Non-communicable diseases contributed 1·73 billion (95% UI 1·54–1·94) DALYs in 2021, with a decrease in age-standardised DALY rates since 2010 of 6·4% (95% UI 3·5–9·5). Between 2010 and 2021, among the 25 leading Level 3 causes, age-standardised DALY rates increased most substantially for anxiety disorders (16·7% [14·0–19·8]), depressive disorders (16·4% [11·9–21·3]), and diabetes (14·0% [10·0–17·4]). Age-standardised DALY rates due to injuries decreased globally by 24·0% (20·7–27·2) between 2010 and 2021, although improvements were not uniform across locations, ages, and sexes. Globally, HALE at birth improved slightly, from 61·3 years (58·6–63·6) in 2010 to 62·2 years (59·4–64·7) in 2021. However, despite this overall increase, HALE decreased by 2·2% (1·6–2·9) between 2019 and 2021. Interpretation: Putting the COVID-19 pandemic in the context of a mutually exclusive and collectively exhaustive list of causes of health loss is crucial to understanding its impact and ensuring that health funding and policy address needs at both local and global levels through cost-effective and evidence-based interventions. A global epidemiological transition remains underway. Our findings suggest that prioritising non-communicable disease prevention and treatment policies, as well as strengthening health systems, continues to be crucially important. The progress on reducing the burden of CMNN diseases must not stall; although global trends are improving, the burden of CMNN diseases remains unacceptably high. Evidence-based interventions will help save the lives of young children and mothers and improve the overall health and economic conditions of societies across the world. Governments and multilateral organisations should prioritise pandemic preparedness planning alongside efforts to reduce the burden of diseases and injuries that will strain resources in the coming decades. Funding: Bill &amp; Melinda Gates Foundation

Interpretation of Rock Mass Behaviour via Multiple Graph Approach: Adit P-C9 of the Alborz Tunnel

The current paper focuses on the application and advantages of the “multiple graph” approach for interpretation of surrounding rock mass behaviour in underground structures. Behaviour of the Argillitic rock mass surrounding Adit P-CP9 of the Alborz Tunnel was interpreted via the “multiple graph” approach resulting in interestingly accurate prediction. The accuracy of the estimation was later observed in the excavation process and afterwards. The observed results are presented which verifies that the “multiple graph” approach can cope satisfactorily with various geological conditions