Alternative approaches for the treatment of Asthma and COPD: Focus on Cell-based therapies, Epigenetics, and Gene silencing approaches

Despite many organized health initiatives and critically acclaimed guidelines for proper management of asthma therapy, there is still a large population of severe asthmatics having an uncontrolled disease. Severe persistent asthma, characterized by chronic airway inflammation, increased eosinophils and serum IgE is currently managed by using inhaled corticosteroids. It is quite challenging to get the best treatment guidelines for bronchial asthma in severe asthmatics, particularly in the presence of steroid resistance and the non-responsiveness to β-agonists. For that purpose, other methodologies are required to reverse the uncontrolled airway remodeling in steroid-resistant severe asthma. These advanced alternative approaches should be able to treat asthma symptoms and to improve the inflammatory conditions underlying characteristic pathological features of asthma. The current review focuses and summarizes the alternative approaches used in severe asthma patients. Agents targeting inflammatory cytokines, phosphodiesterase inhibitors, antibodies, oligonucleotides, stem cells, and target drug delivery using gene silencing, offer promise in treating severe asthma.Keywords: Asthma; COPD; Therapies; Epigenetics; Gene silencin

APOBEC3G Variant (rs6001417) CG and GG Genotypes and their protective feature against HIV-1 Infection in Pakistani Dwelled Community

Background: APOBEC3G (Apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G) gene is one of the genetic host factors, have been linked with HIV-1 AIDS predisposing and protection in different residence populations. The investigation of genetic marker (APOBEC3G) variant (rs6001417) CC, CG and GG genotypes in Pakistan.Methods: The extraction of DNA, the DNA Rapid Salting-out method was used. Then the observed DNA with electrophoresis technique referred for quantitative real-time PCR to identify the APOBEC3G variant rs6001417 genotypes and Taq Man genotyping.  Results: Three genotypes of rs6001417 (CC, CG and GG) were compared both in HIV-1 infected patients and healthy control groups (p=0.73, p=0.007, p=0.01 respectively). The rs6001417 CG and GG genotype demonstrated a significant involvement in both the healthy and infected individuals and portraying possible protective effect against HIV-1 infection with predictive value of 36.43% and 13.57% respectively.Conclusion: APOBEC3G (rs6001417) CG and GG genotypes may have a protective feature in the progression of HIV-1 infection and we may use this as a preliminary predictive marker in the country for HIV-1 infected individuals as well.Keywords: HIV-1; APOBEC3G; Predictive marker; Predictive value; Real-time PC

Male predominant association with Apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G variants (rs6001417, rs35228531, rs8177832) predict protection against HIV-1 infection

Background: Human immunodeficiency virus (HIV) infection, it is a global health concern mainly lead to acquired immune deficiency syndrome (AIDS). There are numerous limitations of this infection particularly in the form of host factors which may limit and interfere HIV-1 replication. The most notable host factors which hinder HIV-1 DNA propagation is the apolipoprotein B mRNA-editing enzyme, catalytic polypeptide- like 3G (APOBEC3G). Any genetic polymorphism of this substantial host factor may impact the host susceptibility pattern to HIV viral infection in different part of the world. The aim of this study to examine genetic variants (rs6001417, rs35228531, rs8177832) effecting HIV-1 infection.Method: Three variants of APOBEC3G gene polymorphism were genotyped while using RT-PCR method. Frequency distribution of these genotypes was evaluated in both the HIV-1 and healthy group.Results: The rs6001417 CG (p = 0.03) and rs35228531 CT (p = 0.01) genotypes were found as protective elements, while rs35228531 TT (p = 0.02) and rs8177832 AA (p = 0.03) genotypes had shown susceptibility against the HIV-1 infection. Our data suggest, rs35228531 CT (p = 0.003) and rs8177832 AA (P = <0.001) genotypes have predominant incidences in HIV-1 male population than healthy control.Conclusion: We predict rs6001417 CG, rs35228531 CT as protective and rs35228531 TT, rs8177832 AA genotypes as a predisposing tool, against the HIV-1 infection in a section of Pakistani population. In addition, male gender was found predominantly high in both protective genotype rs35228531 CT (p = 0.003) and predisposing genotype rs8177832 AA (p = <0.001). The predominant contribution may help the patient to be predict about the status of HIV infection, however, extra efforts are required to study larger cohort of patients to better elucidate the association

Association of anti C1q and ds-DNA levels with the pathogenesis of Lupus Nephritis among SLE patients

Background: Lupus nephritis (LN) is the most common and serious complication associated with SLE and it results in significant morbidity and mortality. It is known by several studies that patients of LN have higher levels of anti-dsDNA and anti-C1q compared with SLE patients without renal involvement. The current study was designed to determine and compare the level of anti-dsDNA and anti-C1q in patients of SLE with and without lupus nephritis in the Pakistani population. This current study was also aimed at providing proof that anti-C1q levels are more prominent in LN/non-LN SLE as compared to anti-dsDNA. This project may help in the determination of results in Pakistan and contribute to the further confirmation of the sensitivity of anti-C1q.Method: The patient samples were collected from Sheikh Zayed hospital, Lahore. These patients were clinically diagnosed by the Rheumatologists as SLE and LN positive on the basis of ACR and SLEDAI scoring criteria. This study was performed and samples were analyzed in the Department of Medical and Laboratory Sciences, Imperial College of Business Study, Lahore on the patient’s serum by ELISA technique.Result: About 38% (12) patients with LN were positive for anti-dsDNA and 31% (9) SLE patients without LN were positive whereas about 38.7% (12) were anti-dsDNA negative in LN cases and 58.6% (17) in SLE without LN. In case of anti-C1q 100% (31) of these LN patients were positive and 93.1% (27) patients SLE without LN showed positive anti C1q results. Only 6.9% (2) patients showed negative results for anti-C1q in LN negative patientsConclusion: The higher levels of anti-C1q suggest that it may be a better diagnostic marker for LN than that of anti-dsDNA and that it can be helpful in the prognosis of SLE patients

Expression level of serum Interleukin-37 in Rheumatoid Arthritis patients and its correlation with Disease Activity Score

Background: Interleukin-37 (IL-37) is a member of IL-1 cytokine family. IL-37 immunosuppresses the pathogenesis of rheumatoid arthritis via down-regulating pro-inflammatory cytokines. The aim of the current study was to evaluate the expression level of IL-37 in rheumatoid arthritis (RA) patients and its correlation with the disease activity score in 28 joints (DAS-28).Methods: In the current study, forty-six RA patients, having a ratio of 19 males and 27 females, and twenty healthy controls (11 males and 9 females) were included. DAS-28 was measured on the basis of patients’ clinical observations of the tender and swollen joints, physical examination and erythrocyte sedimentation rate (ESR). ESR was measured according to the Westergren method. Serum IL-37 level was measured by ELISA. Depending upon the DAS28 calculations the patients were divided in four groups as; 19 in remission, 6 had mild disease activity, 6 were in moderate state and 15 patients were found with severe disease activity.Results: Serum IL-37 levels were found markedly raised in RA patients (mean = 862.6) than in healthy individuals (mean ± SD = 4.4 ± 1.74 pg/ml). Further, our results suggest that level of IL-37 increased significantly from mild (mean ± SD = 829.17 ± 61.40 pg/ml) to moderate (mean ± SD = 1307.5 ± 165.1 pg/ml) and severe (mean ± SD = 1607 ± 86.8 pg/ml) disease prognosis.Conclusion: Thus we conclude, IL-37 has a positive correlation with DAS28 and thus has a potential role in RA pathogenesis. Keywords: Interleukin 37, rheumatoid arthritis, autoimmune disorder, inflammation, disease activity scor

Protective role of ITPA rs1127354-CA polymorphism against anemia in HCV patients using sofosbuvir ribavirin therapy: age and gender match case-control study

Background: Hepatitis C virus is affecting around 80 million people. Sofosbuvir ribavirin-based therapy is associated with certain side effects, especially anemia. Inosine triphosphatase (ITPA) genetic polymorphisms cause functional impairment in ITPase enzyme, leading protection against anemia and improving sustained viral response. This study aims to explore the impact of ITPA variants on hemoglobin decline, ribavirin dose reduction, and sustained viral response (SVR) achievement.Methods: This is prospective gender and age matched case-control study of HCV genotype-3a infected individuals taking sofosbuvir-ribavirin treatment. Patient CBC, viral RNA, liver function test, and ribavirin dose reduction were recorded monthly. ITPApolymorphisms-rs1127354 were determined and confirmed by restriction fragment length polymorphism and sanger sequencing. Effects of polymorphism on hemoglobin level, ribavirin dose and treatment outcome were analyzed.Results: ITPA rs1127354-CC genotype patients experience significant reduction in level of Hb leading to ribavirin dose reduction. Low mean Hb levels were observed in these individuals at first and last month of treatment. No statistical difference was observed in adverse effects on basis of ITPA genotype except fever. Age, BMI, and ITPA genotype rs1127354-CC were independently associated (p< 0.05) with a decrease in Hb level ≥ 2g/dl below the baseline and ribavirin dose reduction. All patients with rs1127354 CA-genotype achieve SVR.Conclusion: Pretreatment determination of ITPA polymorphism can further optimize HCV treatment with new direct-acting antivirals. ITPA rs1127354-CA has a protective role against ribavirin-associated anemia development and individualized management of ribavirin dose and along with the achievement of better SVR rates

CT analysis of subcutaneous and visceral adipose tissue in normal BMI subjects: association with level of physical activity and hypertension

Background: The distribution of adipose tissue, complex factors affecting it and its pathological consequences are among the hot topics in medical research nowadays. Most of the studies reported in the literature however describe the association of factors affecting the fat distribution in overweight and obese individuals. This particular study was however planned to find out the same in subjects having normal basal metabolic index (BMI). The objectives of the study were to analyze total adipose tissue (TAT), subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) in the abdomen volumetrically using CT, to establish the association of these to the levels of physical activity, presence or absence of hypertension and to compare these associations in both the genders.Methods: A prospective study was carried out on seventy five, normal BMI subjects aged between 20–50 years. CT imaging was used for volumetric measurement of TAT, SAT and VAT. Pearson’s correlation of these were then found out with age. Kruskal Wallis test was also performed to compare these in hypertensive and non-hypertensive subjects and in those with different physical activity levels (PAL).Results: Women showed significantly higher volumes of TAT and SAT. Men showed statistically significant correlations of TAT and VAT with age. SAT volumes had significant negative association with the PAL in both genders. Men showed higher responsiveness of fat deposition in all compartments to the presence of hypertension.Conclusion: In conclusion, factors such as gender, age, level of physical activity and hypertension affect the site specific deposition of fat even in those individuals who aren’t over-weight or obese.Keywords: Total Adipose Tissue (TAT); Subcutaneous Adipose Tissue (SAT); Visceral Adipose Tissue (VAT

Temporal and Quantitative Analysis of Atherosclerotic Lesions in Diet-Induced Hypercholesterolemic Rabbits

The diet-induced atherosclerotic rabbit is an ideal model for atherosclerosis study, but temporal changes in atherosclerotic development in hypercholesterolemic rabbits are poorly understood. Japanese white rabbits were fed a high-cholesterol diet to induce sustained hypercholesterolemia, and each group of 10–12 animals was then sacrificed at 6, 12, 16, or 28 weeks. The rabbit aortas were harvested, and the sizes of the gross and intima atherosclerotic lesions were quantified. The cellular component of macrophages (Mφs) and smooth muscle cells (SMCs) in aortic intimal lesions was also quantified by immunohistochemical staining, and the correlation between plasma cholesterol levels and the progress of atherosclerotic lesions was studied. The ultrastructure of the atherosclerotic lesions was observed by transmission electron microscopy (TEM). Widely variable atherosclerotic plaques were found from 6 weeks to 28 weeks, and the lesional progress was closely correlated with cholesterol exposure. Interestingly, a relatively reduced accumulation of Mφ, an increased numbers of SMCs, and a damaged endothelial layer were presented in advanced lesions. Moreover, SMCs were closely correlated with cholesterol exposure and lesional progress for the whole period. Cholesterol exposure directly determines atherosclerotic progress in a rabbit model, and the changes in the cellular component of advanced lesions may affect plaque stability in an atherosclerotic rabbit model