Anti-Mucin1 aptamer-conjugated chitosan nanoparticles for targeted co-delivery of docetaxel and IGF-1R siRNA to SKBR3 metastatic breast cancer cells

Background: Targeted co-delivery of siRNA and a chemotherapeutic drug is an attractive approach to cancer drug design and treatment. This study was carried out to design an anti-Mucin1 aptamer [Apt]-conjugated chitosan nanoparticle [NP] for targeted co-delivery of insulin-like growth factor receptor 1 [IGF-1R] Silencer siRNA and docetaxel [DTX] to SKBR3 cells

Methods: Characterization of nano-drugs, cellular uptake of NPs, cell viability, and gene expression studies were evaluated based on metastatic breast cancer cells

Results:The results of this study showed that NPs had spherical and smooth morphology with 110-118 nm in size and had positive zeta potential [12-14 mV]. siRNA and DTX were considerably loaded into NPs. The appropriate conjugation of the Apt to the NPs was affirmed by gel electrophoresis. The Apt-conjugated NPs were observed to enhance the cellular uptake of NPs into the SKBR3 cells. Although the combination treatment significantly decreased the cell viability of SKBR3 cells, the augmentative effect was observed when Apt was conjugated to NPs. Furthermore, Apt-conjugated NPs dramatically reduced the genetic expression of IGF-1R, signal transducers and activators of transcription 3 [STAT3], matrix metalloproteinases [MMP9], and vascular growth factor [VEGF]

Conclusion: The targeted NPs may augment the targeting of pathways involved in tumorigenesis and metastasis of breast cancer. Therefore, more animal model experiments are needed to further clarify the efficacy and safety of this functionalized nanodrug

Evaluation of the role of TAX, HBZ, and HTLV-1 proviral load on the survival of ATLL patients

BACKGROUND: Adult T-cell leukemia/lymphoma (ATLL) is an aggressive malignancy with very poor prognosis and short survival, caused by the human T-lymphotropic virus type-1 (HTLV-1). The HTLV-1 biomarkers trans-activator x (TAX) and HTLV-1 basic leucine zipper factor (HBZ) are main oncogenes and life-threatening elements. This study aimed to assess the role of the TAX and HBZ genes and HTLV-1 proviral load (PVL) in the survival of patients with ATLL. METHODS: Forty-three HTLV-1-infected individuals, including 18 asymptomatic carriers (AC) and 25 ATLL patients (ATLL), were evaluated between 2011 and 2015. The mRNA expression of TAX and HBZ and the HTLV-1 PVL were measured by quantitative PCR. RESULTS: Significant differences in the mean expression levels of TAX and HBZ were observed between the two study groups (ATLL and AC, P=0.014 and P=0.000, respectively). In addition, the ATLL group showed a significantly higher PVL than AC (P=0.000). There was a significant negative relationship between PVL and survival among all study groups (P=0.047). CONCLUSION: The HTLV-1 PVL and expression of TAX and HBZ were higher in the ATLL group than in the AC group. Moreover, a higher PVL was associated with shorter survival time among all ATLL subjects. Therefore, measurement of PVL, TAX, and HBZ may be beneficial for monitoring and predicting HTLV-1-infection outcomes, and PVL may be useful for prognosis assessment of ATLL patients. This research demonstrates the possible correlation between these virological markers and survival in ATLL patients.

Evaluation and comparison of body mass index and albumin level in patients with active tuberculosis and latent tuberculosis infection

Introduction: Limited data are available on the relationship between nutritional status and tuberculosis. The aim of this study was to evaluate and compare the body mass index [BMI] and serum albumin level in patients with active tuberculosis [ATB] and latent tuberculosis [LTB]

Materials and Methods: A cross-sectional study was conducted on 17 patients newly diagnosed with pulmonary TB who were referred in Iran, during September 2011 to March 2012 and 17 latent tuberculosis infection individuals. Standard method was performed to collect an early morning fasting blood sample for albumin [by the bromocresolgreen method]. Also [BMI] was calculated as body weight divided by height squared [kg/m2]

Results: One-sample Kolmogorov-Smirnov test was used to check normal distribution data The mean +/- Standard deviation[SD] for albumin in the patients and controls were 3.62 +/- 0/56 and 4.68 +/- 0.25, respectively. BMI in the patients and controls were 19.46 +/- 2.79 and 25.4 +/- 3.46, respectively. The serum albumin level was significantly lower in the patient group, compared to the control group [P<0.001].BMI was significantly lower in the patient group, compared to the control group [P<0.001]

Conclusion: Our findings demonstrated that BMI and serum Albumin were significantly lower in the active tuberculosis patients than latent tuberculosis groups

relationship between serum pro-brain natriuretic peptide [Pro-BNP] levels and pulmonary arterial hypertension [PAH] in patients with limited scleroderma

Introduction: Pulmonary arterial hypertension [PAH] is a late progressive sclerodermarelated complication, which can lead to right heart failure and cor pulmonale. Given that cardiac catheterization is a diagnostic method of choice for PAH, and considering the high risks of this method, the purpose of this study was to evaluate the relationship between serum Pro-Brain natriuretic peptide [Pro-BNP] Levels and PAH in patients with limited scleroderma

Materials and Methods: In this cross sectional study, during June 2011- Dec 2013, referring patients to two major educational hospitals, Mashhad- Iran, with scleroderma, who were afflicted with the disease for at least two years [or more], were enrolled in the study if they met the inclusion and exclusion criteria. All the patients underwent echocardiography to determine the pulmonary artery pressure [PAP]. Afterwards, the subjects were referred to a lung center for performing body plethysmography, carbon monoxide diffusing capacity [DLCO], and 6-minute walk test [6MWT]. Pro-BNP Serum level was determined using flourescent immune assay method

Results: The present study included 20 patients [18 female subjects] with the mean age of 43.28 +/- 9.56 yrs, and the mean pro-BNP level of 138 pg/ml. The logarithmic correlation between PAP values, Forced Vital Capacity /DLCO ratio, and pro-BNP level, which was measured using Pearson's correlation coefficient, showed a significant association among these variables[ respectively, r=0.76, P=0.001; r=0.677, P=0.011]. Moreover, the DLCO decrease was associated with increasing pro-BNP level, though this relationship was not significant

Conclusion: This study showed that there was a significant relationship between the serum levels of pro-BNP marker and increased PAP in the echocardiography, DLCO reduction, and FVC/DLCO increase. In fact, this serum marker can be used in patients with systemic scleroderma [SSc] to evaluate the status of PAH

Serum level of trace elements [zinc, lead, and copper], albumin and immunoglobulins in asthmatic children

Bronchial asthma [BA] is a chronic inflammatory disease and it is a major health problem. Trace elements such as copper and zinc are essential components of anti-oxidant enzymes and optimal function of the immune response. Changes in the levels of these elements may lead to increase the risk of asthma. The study group consisted of 175 asthmatic children and 165 control group of healthy general population who attend the outpatient allergic clinic [Bou Ali Hospital] in Sari, Mazandaran, Iran between August 2010 and March 2011. Complete blood count, eosinophil count and serum total IgE level and Serum trace element levels [Zinc, lead and copper] were measured in both groups. There was a significant difference in serum levels of copper, lead, IgE [increased], and decreased IgA, between two groups [p=0.001]. There was no significant difference in blood zinc levels and eosinophilia between two groups [p=0.732 and 0.068, respectively] Increased serum levels of copper and lead may be associated with asthma

Expression of IL-17 and COX2 gene in peripheral blood leukocytes of vitiligo patients

Vitiligo is a pigmentation disorder in which inflammatory mediators such as cytokines have a pivotal role in disease's pathogenesis. Interleukin 17 [IL-17A] is a proinflammatory cytokine which is involved in the induction of several proinflamatory mediators such as cyclooxygenase 2 [COX2]. The aim of this study was to investigate the gene expression of IL-17 and COX2 in peripheral blood leukocytes of vitiligo's patients. Peripheral blood leukocytes from 15 patients with vitiligo and 15 healthy controls were separated using a gradient density centrifugation method. After total RNA isolation and cDNA synthesis, IL-17 and COX2 gene expression were quantified by real-time polymerase chain reaction [PCR]. There were no significant differences in IL-17 and COX2 gene expression in lymphocytes of patients with vitiligo compared with control group [p<0.05]. However there was an increased IL-17 and COX2 gene expression in neutrophils of patients compared to controls, but it did not reach statistical significance [p=0.05]. We could not find any differences in IL-17 and Cox2 gene expression between clinical diseases subtypes, sex and age. There was a significant correlation between IL-17 and COX2 genes expression in the neutrophils of patients [p=0.00, r=0.80]. Our results showed an increased expression in IL-17 and Cox-2 genes in neurophils of patients with vitiligo. This suggested that these two factors are involved in the inflammatory process. Further studies with a larger sample size might help to establish the role of these factors in the pathogenesis of diseases

[Skin prick test in asthmatic, allergic rhinitis and urticaria patients]

Asthma, allergic rhinitis and urticaria are common diseases in the world. The aim of this study was to investigate the aeroallergen by skin prick test in patients, in Mazandaran province, Sari [Iran]. This was a prospective study of skin prick test of aeroallergens in patients with asthma, allergic rhinitis or both and urticaria based on clinical and Para clinical criteria. Eight hundred and nine cases were referred to Tooba and Boo-Ali allergic centers in Mazandaran University of Medical Sciences between December 2006 and July 2010. The aeroallergens were included; House dust mites [DF, DP], Coach Roach, Feather, Aspergillus, Alternaria, Pigweed, Nettle, Oak, Maple and others. Negative [normal saline] and positive controls [histamine] were applied for all patients. A total of three hundred and fifty out of 809 cases were males [43%] and 459 cases were Females [57%]. The age range was between 5 and 50 years old, 42.5 percent [n=340] of patients had ages between 10 and 20 years old. In asthma, allergic rhinitis or both and urticaria, the prick test DF 32%, DP 33%, Cockroach 22% and feather 9 were positive. Other allergens were positive between 0 and 10 percent, except candida which was 28% positive. Total IgE levels and Eosinophil count was increased in 54 and 35 percent of cases, respectively. Due to warm and humid weather in north of Iran [Sari], house dust mites are more common

functional polymorphism of the granulocyte macrophage colony stimulating factor is not associated with the outcome of HTLV-I infection

Genetic background has known to be associated with the outcome of human T cell lymphotropic virus [HTLV] type I infection. In The present study we investigate the association between GM-CSF gene polymorphisms with the outcome of HTLV-I infection. We analyzed 3 single-nucleotide polymorphisms in the promter region of granulocyte macrophage colony stimulating factor [GM-CSF] at positions -677A/C, -1440A/G and -1916T/C in 68 patients with HTLV- I-associated myelopathy/tropical spastic paraparesis [HAM/TSP] and 77 HTLV-I-seropositive asymptomatic carriers and 175 healthy controls from an area in Iran, Mashhad, where HTLV-I is endemic. No significant differences were observed in the distribution of GM-CSF polymorphisms between HAM/TSP patients, HTLV-I carriers and healthy controls [P> 0.05]. The -611A/C polymorphism fall within the transcriptional enhancer factor-2 [TEF-2] binding site, so an electrophoretic mobility shift assay [EMSA] was performed to determine the effects of polymorphisms on protein binding to the GM-CSF promoter. The result showed a significantly higher binding efficiency of nuclear protein to the A allele compared with the C allele. Our study suggests that polymorphisms in the GM-CSF promoter is not associated with the outcome of HTLV-I infection, however, GM-CSF polymorphism at position -677 could indeed influence gene expression

Association between the polymorphisms of IL-4 gene promoter [-590C>T], IL-13 coding region [R130Q] and IL-16 gene promoter [-295T>C] and allergic asthma

Allergic asthma is a multifactorial disease, influenced by genetic and environmental factors. Recent family-based studies have revealed evidence for linkage of human chromosomes 5q31-33, 12ql5-24, Ilql3 and 15q23.6 as regions likely to contain genes related to asthma. Among the candidate genes in these regions are the genes encoding for human interleukin-4, interleukin-13 and interleukin-16. To evaluate this linkage, we examined an Iranian population of patients with asthma. A total of 30 patients with allergic asthma and 50 normal subjects were studied. Allergic asthma was confirmed using skin prick test and spirometry. DNA was extracted from blood cells and IL-4 [-590C>T], IL-13 [R130Q] and IL-16 [-295T>C] polymorphisms were determined by PCR-RFLP method. Out of 30 patients with allergic asthma, the following genotypes for IL-4, IL-13 and IL-16 cytokines were found: IL-4 genotypes consisted of 17 [56.7%] CC, 8 [26.7%] CT and 5 [16.7%] TT; IL-13 genotypes consisted of 11 [36.7%] GG, 13 [43.3%] GA and 6 [20%] AA; IL-16 genotypes consisted of 23 [76.7%] TT and 7 [23.3%] CT. No patient showed CC genotype for IL-16. A higher proportion of case subjects with the C allele for the IL-4, G allele for the IL-13 and T allele for the IL-16 polymorphisms was found compared with the T, A and C alleles, respectively. These results suggest an influence of genetic variability at the promoter of IL-4 gene [-590C>T] and a coding region of IL-13 gene [R130Q] on the occurrence of allergic asthma and no relationship between IL-16 promoter polymorphism [-295T>C] and this disease

Association between HLA-DRB1 *01 and HLA-Cw*08 and outcome following HTLV-I infection

Human T cell lymphotropic virus type I [HTLV-I]-associated rnyelopa-thy/tropical spastic paraparesis [HAM/TSP] is an inflammatory disease which occurs in less than 2% of HTLV-I -infected individuals. High proviral load, high HTLV-I-specific CD8[+] cytotoxic T lymphocyte frequency [CTL] and host genetic factors such as HLA all appear to be associated with HTLV-I infection. Previous studies have shown that HLA-DRB1*01 increases the risk of HAM/TSP in Japanese HTLV-1 infected individuals. To investigate the association between HLA class II DRB1 alleles and HLA class I alleles [HLA-Cw*08, B54, A*02 and A-30] in HTLV-I infected individuals in Mashhad. Here we determined the frequency of HLA class II DRBl, using INNO-LIPA reverse hybridization line probe assay, and HLA class I alleles [HLA-Cw*08,B54, A*02 and A-30] by PCR-SSCP method in healthy controls, HAM/TSP patients and HTLV-I infected individuals born and resident in Mashhad. The frequency of HLA-DRB1*01 alleles in this population was different from other areas of Iran. The frequency of HLA-DRB1*01 was significantly increased in HAM/TSP patients compared with carriers [p 0.028; OR=9.4]. The frequency of HLA-Cw*08 was also significantly increased in HAM/TSP patients compared with controls [p=0.03; OR=13.5]. Our results may suggest that possession of HLA-DRB1*01 increases the risk of HAM/TSP in HTLV-I-infected individuals and HLA-Cw*08 correlates with low CTL immune response in HAM/TSP patients

HTLV-I infection: virus structure, immune response to the virus and genetic association studies in HTLV-I-infected individuals

Although the structure of human T lymphoptropic virus type I [HTLV-I] has been known well, the function of some proteins encoded by HTLV-I PX region is not fully understood. Furthermore, the responses of the immune system to HTLV-I remain still unknown. Most of HTLV-I infected individuals show a strong and persistently activated cytotoxic T-cell [CTL] response to the virus. The frequency of HTLV-I specific CTL is higher in patients with HTLV-I -associated myelopathy/ tropical spastic paraparesis [HAM/TSP] compared with HTLV-I carriers. However, the efficacy of the immune response determines the outcome of HTLV-I-associated diseases. Among the risk factors which contribute to the observed differences between HAM/TSP patients and HTLV-I carriers, the interaction between different genes and/or environmental factors seem to be important. These factors may also involve in outcome of HTLV-I infection in infected- individuals

Association between the polymorphism of TGF- beta 1 gene promoter [-509C>T] and idiopathic chronic Urticaria

Idiopathic Chronic Urticaria [ICU], the most common form [70-80%] of chronic urticaria is supposed to have immune basis causes. It is speculated that the promoter polymorphism of TGF- beta 1 gene may be involved in ICU. This condition is thought to affect at least 0.1% of the population and often can be severe and difficult to treat. A total of 40 patients with ICU and 41 normal subjects were studied. DNA was extracted from whole blood and TGF- beta 1 promoter -509C>T polymorphism was determined by PCR-RFLP method. Out of the 40 patients with ICU, 11 [27.5%] had CC, 26 [65%] had CT and 3 [7.5%] had TT genotypes. A higher proportion of case subjects with the C allele [CT type or CC type] was found compared with the T allele. These results do suggest an influence of genetic variability at the promoter of TGF- beta 1 gene [-509C>T] on the occurrence of ICU. This polymorphism has been shown as a useful genetic change in our study. Further work is required to confirm this result