Genome editing from bench to clinic by CRISPR/Cas9

Background: Engineered molecular scissors invention beside their ability to conduct site-specific modification of the genome hold great promise for effective functional analyses of genes, genomes and epigenomes. These technologies could improve researchers' understanding of the molecular underpinnings of disease states and facilitate novel medical genetic therapeutic applications. The CRISPR/Cas9 system's simplicity, facile engineering and amenability to multiplexing make it the system of choice for many applications. CRISPR/Cas9 has been used to generate disease models to study genetic diseases. It is expected that in the near future researchers will refinement this technology in various aspects of the CRISPR/Cas9 system, including the system's precision, delivery and control over the outcome of the on-targrt activity during repair process. Here, we discuss the CRISPR/Cas9 history, type of CRISPR systems, the status of this technology in the field of genome editing and its use in new treatments for diseases such as genetics, cancer, etc

Lithophagia as a clue for celiac disease: a case report and literature review

Lithophagia is a type of pica that might be resulted from Iron Deficiency Anemia [IDA]which is the frequent presenting signs of Celiac Disease [CD]. A 5-year-old child with a two year history of the lithophagia with a, refractory IDA, abdominal distention and constipation. The child did not grow well and had failure to thrive. With suspicion to CD, TTg IgA level was measured and due to an incearse of TTg IgA level the patients were undergone esophagogastrodeudonoscpy and jejunal biopsy. The biopsy showed severe villous atrophy and an increase in limphoplasma cells. Biopsy confirmed diagnosis of CD and glutten free diet was initiated finally. Six months after diagnosis and commencing the gluten free diet, the lithophagia and constipation in patient eradicated completely. IDA and failure to thrive were improved and the level of TTg IgA was reached to the normal. The case demonstrated the relationship between lithophagia and CD in anemia. Therefore, in the same cases such as our case should be considered CD as the most important causes of lithophagia

Serological evaluation of celiac disease in children with congenital heart defect; a case control study

Due to the increased prevalence of celiac disease in chromosomal anomalies and other congenital anomalies, this study was conducted to evaluate the seroprevalence of celiac disease [CD] in patients with congenital heart defects [CHD]. This case-control study was done on 1002 children in two groups of CHD patients [n=402] and controls [n=600]. The serum tissue transglutamianse [TTG] levels were investigated. The two groups were compared in terms of TTG IgA levels and p<0.05 was considered as the significant level. The means of serum TTG IgA levels in children with CHD and the control groups were 19.17 +/- 46.67 and 7.77 +/- 10.02 u/mL respectively [p=0.001]. After ANOVA analysis a significant difference between two cyanotic and acyanotic subgroups of cases and control groups was observed [p=0.000]. The follow up tukey test showed only non-significant difference between the cyanotic and acyanotic cases. The frequency of TTG IgA with the consideration of 20 u/mL as cut-off point showed a significant association with groups [X2=28.31 and p=0.000]. According to the results the serum TTG IgA levels were significantly higher in patients with CHD than normal children and screening for CD in children with CHD is recommended

Serologic evaluation of celiac disease in patients with beta thalassemia major and control

This study reports evaluated prevalence of CD in patients with Beta-thalassemia major. Celiac Disease [CD] is an autoimmune disorder triggered by ingestion of gluten in genetically predisposed individuals. In this case-control study in a period of 3 years, which was performed on 620 children in two groups of Beta-thalassemia major patients [n=200] and control [n=420], serum tissue transglutamianse [tTG] IgA levels were measured. The two groups were compared together in terms of tTG IgA levels, and p<0.05 was considered significant. The means of serum tTG IgA levels in patients with Beta-thalassemia major and control groups were 28.81+/-68.44 and 6.94+/-6.68 U/mL, respectively. There was a significant difference in favor of the case group [p=0.000]. Body mass index in the two case and control groups had a significant difference [t=3.859, p=0.001]. Belonging to each group will change the probability of having less than 20 in tTG IgA [odds=0.285] and it means that belonging to the control group has a protective role. There is only a significant association in the case of all population [r=0.102, p=0.011]. Body mass index in the two case and control groups had a significant difference [t=3.859, p=0.001]. Probability of CD should be considered since the prevalence of CD is high in patients with and Beta-thalassemia major. Patients with thalassemia major are recommended for screening for CD

Combination of erythromycin and propranolol for treatment of childhood cyclic vomiting syndrome: a novel regimen

This study aimed to evaluate the erythromycin efficacy in childhood cyclic vomiting syndrome. Cyclic vomiting syndrome [CVS] is an unusual cause of episodic emesis in children and erythromycin is an effective treatment. In this prospective study, 301 patients with a final diagnosis of CVS enrolled in two separated groups. The first group received erythromycin for 7 days and propranolol for at least 9 months [n=155]. The second group was treated with propranolol alone for at least 9 months [n=146]. These two groups were compared for response to the treatment and the recurrence of symptoms after treatment completion. Relationship of response, recurrence, and characteristics of the disease was assessed. Both groups showed a significant difference in terms of response to treatment [P=0.002], however the recurrence after treatment completion had no considerable difference [P=0.563]. There was no relationship between CVS characteristics and these two items [response and recurrence]. In our point of view, the addition of erythromycin to standard propranolol treatment can improve the response to treatment, although it has no significant effect on recurrence of CVS symptoms. We suggest the use of erythromycin for 7 days in addition to CVS standard therapy

Mortality risk prediction by application of pediatric risk of mortality scoring system in pediatric intensive care unit

The Pediatric Risk of Mortality [PRISM] score is one of the scores used by many pediatricians for prediction of the mortality risk in the pediatric intensive care unit [PICIT]. Herein, we intend to evaluate the efficacy of PRISM score in prediction of mortality rate in PICU. In this cohort study, 221 children admitted during an 18-month period to PICU, were enrolled. PRISM score and mortality risk were calculated. Follow up was noted as death or discharge. Results were analyzed by Kaplan-Meier curve, ROC curve, Log Rank [Mantel-Cox], Logistic regression model using SPSS 15. Totally, 57% of the patients were males. Forty seven patients died during the study period. The PRISM score was 0-10 in 71%, 11-20 in 20.4% and 21-30 in 8.6%. PRISM score showed an increase of mortality from 10.2% in 0-10 score patients to 73.8% in 21-30 score ones. The survival time significantly decreased as PRISM score increased [P<0.001]. A 7.2 fold mortality risk was present in patients with score 21-30 compared with score 0-10. ROC curve analysis for mortality according to PRISM score showed an under curve area of 80.3%.PRISM score is a good predictor for evaluation of mortality risk in PICU

relationship between personality traits, anxiety and depression, in life quality of patients under treatment by haemodialysis [HD]

The purpose of this study is to examine personality traits in renal patients undergoing haemodialysis [HD] and its connection to depression, anxiety, and life quality. In this study we also aim to propose an intervention for treatment and prevention of these personality traits. This cross-sectional study was performed on 70 patients affected by chronic renal failure undergoing haemodialysis treatment. Patients were selected based on an accessible sample. Required data were gathered through questioners: Hospital Anxiety and Depression, NEO-FFL, and SF-36 Health Survey. Based on the results obtained from this study 47.1% of patients undergoing HD treatment fall into the abnormal category according to the rate of depression. There is a positive meaningful correlation between conscientiousness and agreeableness and a negative correlation between extroversion, anxiety, and depression in life quality of renal patients. With respect to the correlation existing between the quality of life and the personality traits, anxiety and depression, in patients under HD treatment, life quality and treatment outcome can be improved if psychological problems are diagnosed early and psycho education and social interventions are presented

[Cancer metastasis, genetic and microenvironmental factors of distant tissue: a review article]

Cancer is one of the main reasons of mortality worldwide, and more than 90 percent of cancer deaths are due to metastasis. Although primary tumors are curable using chemical adjuvant therapy or surgery, metastatic tumors are mostly incurable. This resistance shows the high rate of mortality among patients with metastatic disease. Being a sequential event, metastasis is a subtle and intricate process in which tumor cells undergo a plenty of changes and acquire the capacity of migration, invasion, survival and self-renewal which all are necessary for metastasis to happen. The key point in recognition and cure in invasive cancers is to identify critical genes, proteins and pathways involved, and show their relation with each other and the disease. Forming metastasis needs favorable genetic and microenvironmental elements of tumor cells and distant tissue, respectively. Unfavorable conditions in each steps of this process lead to arresting metastasis and subsequent dormancy, which is the most important phenomenon in relapse. In this review, benefiting from tens of reliable and recently identified data and personal experiences, it has been tried to draw new patterns associated with metastasis for further investigation. Determining genes, proteins and microenvironmental factors that affect metastasis, in a sequential manner, can help better understanding of this lethal process and subsequently a prosperous treatment

Correlation between PPAR gamma protein expression level in granulosa cells and pregnancy rate in IVF program

Peroxisome proliferative-activated receptors [PPARs] are nuclear receptors that involved in cellular lipid metabolism and differentiation. The subtype gamma of the PPAR family [PPAR gamma] plays important roles in physiologic functions of ovaries. To determine correlation between PPAR gamma protein level in granulosa cells and pregnancy rate in women undergoing in-vitro fertilization [IVF] treatment. In this cross-sectional study, twenty-five samples of granulosa cells were collected from women referred to an IVF treatment center. PPAR gamma protein expression level in granulosa cells was determined in comparison with beta -actin level as control gene with Western blot test. Laboratory pregnancy was determined by a rise in blood beta -hCG level fourteen days after embryo transfer. Correlation analyses were used to test for associations between the oocytes and pregnancy occurrence as outcome variables and PPAR gamma protein expression level. Correlation analysis indicated that there was no significant relationship between granulosa cells PPAR gamma protein level with IVF parameters including number of matured oocytes and the ratio of fertilized to matured oocytes. Comparison of granulosa cells PPAR gamma protein level with positive and negative laboratory pregnancy revealed also no significant relationship. According to the results of this study, PPAR gamma protein level in granulosa cells could not be directly correlated to the success rate of IVF

Monocyte expression of toll-like receptor-4 in patients with stable angina undergoing percutanoeus coronary intervention

Toll like receptors [TLRs] are well recognized players in inflammatory conditions. Among them TLR-4 is involved in chronic inflammatory processes such as formation of atherosclerotic plaques. The present study was aimed to examine the effects of percutanoeus coronary intervention [PCI] as a revascularization method on monocyte expression of hTLR-4 and on the serum levels of two proinflammatory cytokines [TNF-alpha and IL-1beta]. Blood samples were obtained from 41 patients with stable angina who were candidates for PCI. The samples were collected immediately before and 2h and 4h after PCI. The expression of hTLR-4 on CD14[+] monocytes and the serum levels of TNF-alpha and IL-1beta were measured using flowcytometry and ELISA techniques, respectively. By comparing the frequency of circulating hTLR-4[+]/CD14[+] monocytes at different time points, it was observed that PCI procedure up regulates the monocyte expression of hTLR-4 [p<0.05]. The increase in expression was associated with the elevation of the serum levels of proinflammatory cytokines [p<0.05]. There was a significant correlation between monocyte expression of hTLR-4 and serum levels of TNF-alpha and IL-1beta only before PCI. In spite of parallel increase in the serum levels of proinflammatory cytokines and the monocyte expression of hTLR-4, the correlation did not attain a significant level after PCI intervals. PCI is positively associated with an increase in the monocyte expression of hTLR-4. It is also associated with the elevation in the serum levels of proinflmmatory cytokines. These findings suggest that hTLR-4 monocyte expression may be used as a potential prognostic tool in patients with stable angina undergoing PCI.

[Roles of miRNAs in gene expression regulation, apoptosis, diagnosis and treatment of cancer]

Apoptosis, as a programmed cell death, is activated in response to various stresses. Apoptosis is an active and energy need process that many mechanisms and genetic factors control it with specific programs. Apoptotic cells have specific features. Apoptosis acts in many different organic evolutionary stages. Aberrations in regulation of this process can cause many diseases, such as cancers. Different molecules, like miRNAs, have a role in apoptosis process. miRNAs, as regulatory molecules, act in abundant cellular mechanisms. Regarding their function, apoptotic related miRNAs were divided in proapoptotic and antiapoptotic groups. miRNAs target several mRNAs and also themselves were regulated by other genes. This regulatory loop between miRNAs and related genes cause complexity. miRNAs have different expression profile in normal and abnormal cells. Apoptosis and miRNA both act in different processes of tumorigenesis, metastasis and antiogenesis. miRNAs can be used as biomarkers in prevention, prognosis and prevention of different cancers. Advances in knowledge about miRNAs lead to develop new therapeutic trend. Recent researches have suggested using miRNAs involved in apoptosis in the treatment of cancers

Serum uric acid correlation with echocardiographic indices in children with dilated cardiomyopathy

Dilated cardiomyopathy [DCMP] is a myocardial disease characterized by dilated left ventricle or both, ventricles and reduced contractility of the myocardium. In patients suffering from DCMP, the serum level of uric acid may increase. This research was designed to evaluate the effect of the serum level of uric acid on systolic and diastolic functions in patients with DCMP. This case-control study was performed on 30 patients with DCMP aged between 1 month and 12 years who were consistent with a control group in terms of age and gender. Patients suffering from congenital and acquired cardiac, renal, metabolic, endocrine, musculoskeletal, neurologic, vascular, and hematologic diseases were excluded. After physical examination, chest X-ray, and electrocardiography, systolic and diastolic parameters were measured via echocardiography, and fasting serum uric acid level was measured. The data were analyzed using the t-test and Pearson correlation coefficient. The average age of the patients in the case and control groups was 7.28 and 7.13 years, respectively. There were 15 boys, and the rest were girls. The serum uric acid level in the case and control groups was 6.22 and 3.31 mg/dl, respectively; the difference was statistically significant [P value < 0.01]. There was a significant correlation between serum uric acid level and left ventricular isovolumic contraction, interventricular septal diameter, left ventricular septal diameter in diastole, and fractional shortening [P value < 0.05]. In children with DCMP, the serum level of uric acid increases significantly and this increase is significantly correlated with some of left heart echocardiographic parameters. This test is of predictive value for disease progression

I405V and -629C/A polymorphisms of the cholesteryl ester transfer protein gene in patients with coronary artery disease

Cholesteryl ester transfer protein [CETP] plays a main role in high-density lipoprotein metabolism. CETP gene possesses several single nucleotide polymorphisms which have been associated with plasma high-density lipoprotein cholesterol [HDL-C] concentrations. The aim of this study was to determine the association of CETP -629C/A and I405V polymorphisms with coronary artery disease [CAD] in Iranian population. The presence of two CETP gene polymorphisms -629C/A and I405V were studied in 187 unrelated CAD cases and 136 controls. All the samples were clinically examined and lipid profile was estimated. Genotyping was performed using polymerase chain reaction/restriction fragment length polymorphism method. The frequency of-629C/A and I405V allelic variants were found to be 0.732 and 0.366 in cases and 0.658 and 0.348 in controls, respectively. The frequency of A allele of -629C/A polymorphism in cases was significantly higher than that of controls. HDL-C in AA genotype was higher than CA and CC genotypes in controls. No significant effect of II, IV and VV genotypes was found in lipid profiles. No significant association was found between CETP I405V polymorphism and increased risk of CAD in Azeri population studied. AA genotype of -629C/A increased HDL but the risk of CAD in this genotype might be higher than CC genotype