Dual pH- and temperature-responsive poly(dimethylaminoethyl methacrylate)-coated mesoporous silica nanoparticles as a smart drug delivery system.

A robust drug delivery system was created by grafting poly(dimethylaminoethyl methacrylate) (PDMAEMA) onto silica nanoparticles with two different lengths using an in situ atom transfer radical polymerization, resulting in the formation of a pH- and temperature-sensitive shell. The high molecular weight PDMAEMA demonstrated effective controlled drug release, and prevented drug release in healthy cells. Drug release occurred through polymer shell protonation at pH 5. The critical temperature of 41 °C facilitated rapid solvation of the shell polymers in the blood, preventing tissue accumulation and reducing toxicity compared to systems with lower critical solution temperatures. Field-emission scanning electron microscopy analysis and nitrogen adsorption/desorption analysis showed that the nanoparticles have a fine network, mesoporous structure, and a mean size of around 17 nm that show their excellent capacity for loading drugs. Fourier-transform infrared spectroscopy showed that all the modification steps and polymerization were successfully implemented. Thermogravimetric analysis showed PDMAEMA chains with two different lengths grafted onto the nanoparticles. Transmission electron microscopy analysis also showed grafted polymer chains on the hybrid nanoparticles. The release profile of model cancer drugs (doxorubicin and methotrexate) varied with pH and temperature, with high molecular weight PDMAEMA shells effectively preventing drug release at neutral pH. In vitro analysis using the HeLa cell line showed minimal toxicity in blank samples and significant release profile in acidic environment.

RUNX1 variant as a genetic predisposition factor for acute myeloid leukemia.

Acute myeloid leukemia (AML) is the most common hematological malignancy among adults and is characterized by accumulation of immature myeloid cells. Different genetic factors have role in the occurrence of AML. Among different proteins, RUNX1 and BAALC are involved in the development AML. It has been shown that BAALC overexpression is a factor that indicate shorter disease free survival in a subset of AML patients. RUNX1 has been implicated in the development of breast, prostate, lung, and skin cancers. The aim of this study is determination of the prevalence of common polymorphisms in BAALC (rs6999622 and rs62527607) and RUNX1 (rs13051066 and rs61750222) in AML patients compared with healthy subjects. A total of 100 AML patients and 100 healthy control subjects were included in our study. Genomic DNA was isolated from peripheral blood and the polymorphisms were genotyped by applying ARMS and PCR-RFLP methods. Finally, data was analyzed using SPPSS software. Our results demonstrate a significant association between the RUNX1 rs13051066 and AML in the co-dominant (odd ratio = 6.66, 95% Cl = 1.85-25, p = .006) and dominant (GT + TT versus GG: odd ratio = 6.15, 95% CI = 1.73-21.87, p = .002) models. The RUNX1 rs13051066 polymorphism is associated with risk of AML in Iranian population. Future studies should consider larger sample size for assessment of RUNX1 gene polymorphisms, and employ cytogenetic and molecular analyses in AML patients from different ethnic origins.

Dual biomarkers long non-coding RNA GAS5 and its target, NR3C1, contribute to acute myeloid leukemia.

Acute myeloid leukemia (AML) is a complex hematological neoplasm with poor prognosis. At present, overwhelming evidence indicates that different genetic abnormalities are relevant to the pathogenesis of AML. Nevertheless, its exact molecular mechanism is still unknown. Recently, it was reported that lncRNAs play crucial roles in tumorigenesis. But, their role in the molecular pathogenesis of AML has not been extensively explored. GAS5, one of the earliest known lncRNAs, has an essential role in the formation and progression of multiple human cancers. It was recently demonstrated that GAS5 acts as a riborepressor of the Glucocorticoid receptor) GR) and abnormal levels of GAS5 may alter response of hematopoietic cells to glucocorticoids. GAS5 can have interaction with the GR that encoded by NR3C1 gene and inhibit its transcriptional activity. To test whether the genetic variants can be associated with AML risk, we genotyped rs55829688 (T > C) polymorphism in GAS5 and three NR3C1 SNPs namely rs6195, rs41423247 and rs6189/rs6190 in a population of 100 Iranian AML patients and 100 healthy subjects. The analysis of the data showed the frequency of alleles and genotypes of rs55829688 and rs6189/rs6190 polymorphisms did not differ between patients and healthy subjects. But, rs41423247 and rs6195 demonstrated a significant correlation with AML risk. The rs6195 was associated with higher AML susceptibility in the co-dominant (OR = 4.58, 95% CI = 2.11-9.981, P < .0001), dominant (OR = 4.55, 95% CI = 2.155-9.613, P < .0001), and over-dominant (OR = 4.43, 95% CI = 2.042-9.621, P < .0001) models. Also, the rs41423247 polymorphism was associated with higher risk of AML in co-dominant (OR = 2.07, 95% CI = 1.171-4.242, P = .012) and dominant (OR = 2.47, 95% CI = 1.192-5.142, P = .010) models. Furthermore, haplotype analysis (rs41423247, rs6189.rs6190, rs6195, and rs55829688 respectively) demonstrated that GGAT, CGGT, and GGGT haplotypes were associated with higher risk of AML in the studied population (p-values = .007, 0.042 and 0.044, respectively). The present study reveals a possible role for NR3C1 in the pathogenesis of AML.

Molecular Phylodiagnosis of Enterocytozoon bieneusi and Encephalitozoon intestinalis in Children with Cancer: Microsporidia in Malignancies as an Emerging Opportunistic Infection.

BACKGROUND : Microsporidia may cause infection in both immunocompromised and immunocompetent populations. The best strategy to control microsporidiosis is obtaining thorough knowledge of its outbreak and pathogenicity. PURPOSE : Because of the lack of precise estimation of microsporidia prevalence among Iranian children with cancer, the current study aimed at evaluating the rate of intestinal microsporidia in children undergoing chemotherapy. METHODS:  Patients with cancer undergoing chemotherapy in a children's hospital in Northwestern Iran were studied; 132 stool samples were collected and stained by the Weber and Ryan-blue modified trichrome staining techniques. The extracted DNA samples were evaluated by the nested polymerase chain reaction (PCR) method. All positive isolates were sequenced for genotyping and phylogenetic analysis. RESULTS: A total of 17 (12.8%) samples were microscopically positive for microsporidia infection, whereas only 14 (10.6%) cases were positive based on nested PCR results. In the positive samples detected with nested PCR, the frequency of Enterocytozoon bieneusi and Encephalitozoon intestinalis infections was 71.4% (n = 10) and 28.6% (n = 4), respectively. After sequencing and phylogenetic analysis, the genotype of E. bieneusi was type D and the sequences of the isolated species were similar to those of the registered ones. CONCLUSION: E. bieneusi is a major contributor to microsporidiosis in young immunocompromised patients in Iran. Microsporidia species are well-detected when confirmatory techniques such as molecular methods are in agreement with staining. So, to ensure this, a suggestion has been made to introduce a certain diagnostic test for microsporidiosis.

A Novel Mutation in SNX10 Gene Causes Malignant Infantile Osteopetrosis.

BACKGROUND: Osteopetrosis is a group of genetically heterogonous diseases and the main feature of that is increased bone density due to osteoclast's abnormality. It has three clinical forms based on inheritance pattern, severity and age of onset: the dominant benign form (ADO), the intermediate form (IRO) and the recessive severe form (ARO). One of the recently discovered genes for ARO form is SNX10 that accounts for 4% of affected persons by this type. METHODS: In this paper, a 15 years old girl affected by osteopetrosis has been analyzed for detecting causal mutation in known osteopetrosis genes. To get it done, amplified exons of the genes were sequenced and then were analyzed. RESULTS: Direct sequencing of SNX10 gene showed a homozygous c.43delG variant in the patient. Both healthy parents were heterozygous for this variant. In silico analysis revealed that this novel variant can be considered as the cause of disease in the patient. CONCLUSION: In this paper, a girl affected by osteopetrosis with a novel deletion in SNX10 gene was reported.

Molecular analysis of more than 140 gene fusion variants and aberrant activation of EVI1 and TLX1 in hematological malignancies.

Gene fusions are observed in abnormal chromosomal rearrangements such as translocations in hematopoietic malignancies, especially leukemia subtypes. Hence, it is critical to obtain correct information about these rearrangements in order to apply proper treatment techniques. To identify abnormal molecular changes in patients with leukemia, we developed a multiplex reverse transcriptase polymerase chain reaction (MRT-PCR) protocol and investigated more than 140 gene fusions resulting from variations of 29 prevalent chromosomal rearrangements along with EVI1 and TLX1 oncogenic expression in the presence of optimized primers. The potential of the MRT-PCR method was approved by evaluating the available cell lines as positive control and confirmed by sequencing. Samples from 53 patients afflicted with hematopoiesis malignancies were analyzed. Results revealed at least one chromosomal rearrangement in 69% of acute myeloid leukemia subjects, 64% of acute lymphoblastic leukemia subjects, and 81% of chronic myeloid leukemia subjects, as well as a subject with hypereosinophilic syndrome. Also, five novel fusion variants were detected. Results of this study also showed that chromosomal rearrangements, both alone and in conjunction with other rearrangements, are involved in leukemogenesis. Moreover, it was found that EVI1 is a suitable hallmark for hematopoietic malignancies.

Cryptosporidium infection in children with cancer undergoing chemotherapy: how important is the prevention of opportunistic parasitic infections in patients with malignancies?

Cryptosporidiosis is a relatively uncommon disease in healthy individuals but could be potentially worrisome in immunocompromised patients. This study aimed to evaluate Cryptosporidium infection in children with cancer undergoing chemotherapy. A case-control study was conducted in 132 children with cancer undergoing chemotherapy and 132 non-cancer controls. The modified Ziehl-Neelsen (MZN) staining and polymerase chain reaction methods were used for the detection of Cryptosporidium parasite. All positive isolates were sequenced for phylogenetic analysis. Statistical analysis was performed using the SPSS version 16 and Fisher exact test. The rate of cryptosporidiosis in children with cancer undergoing chemotherapy was 3.8%, which was higher than that of the control group. Other intestinal parasites detected in patients with cancer included Giardia lamblia (3%), Entamoeba coli (1.5%), and Chilomastix mesnili (0.8%). In the control group, only two (1.5%) cases were positive for G. lamblia. No significant difference was observed between the gender, age, residency, contact with domestic animals, stool appearance, neutropenia, chemotherapy period, and type of malignancy with regard to cryptosporidiosis. Phylogenetic analysis revealed that Cryptosporidium parvum isolates in this study relied on a branch that represents similar sequences from Iran and other countries. Although the rate of Cryptosporidium infection was relatively higher in children with cancer undergoing chemotherapy compared to the control group, any statistically significant difference has not been found between them. These findings should not be contrary to the need for healthcare to prevent opportunistic parasitic infections in malignant and immunocompromised patients.

Microsporidiosis in Iran: A systematic review and meta-analysis.

OBJECTIVE: To examine all evidence about Microsporidia infection in vertebrate/invertebrate hosts and Iranian populations distributed in different regions of the country. METHODS: All published articles up to December 2015, including descriptive and cross-sectional studies related to the prevalence and genotyping of Microsporidia infection in Iran, was considered in this systematic review. The meta-analysis was done using the random-effects model and Stats Direct statistical software. MEGA 5.05 software and maximum likelihood algorithm with Kimura 2-parameter model were used for phylogenetic analysis. RESULTS: Of the 1152 investigated studies, 33 eligible studies reported a prevalence of Microsporidia infection in vertebrate and invertebrate hosts. According to this systematic review, the overall prevalence rate of Microsporidia infection in immunocompromised patients in Iran was 8.18%. Furthermore, the overall prevalence rate of Microsporidia infection in immunocompromised patients with chronic diarrhoea, patients with non-diarrhoea, gastroenteritis, and patients with CD4 (<200 cells/µL) was 15.4%, 4.1%, 0.5%, and 12.9% respectively. The highest prevalence rate of human and animal Microsporidia was estimated in Kerman (29%) and Khuzestan (26.5%). The overall prevalence rate of Microsporidia infection in honeybees using the random-effects model was 40%. Furthermore, the highest prevalence rate of nosemosis was described in East Azerbaijan (48.2%). The most Microsporidia isolates from immunocompromised patients and pigeons in Iran belonged to genotypes D (n = 16; 50%) and E (n = 6; 20.6%) of Enterocytozoon bieneusi. CONCLUSIONS: This study may be the first systematic review and meta-analysis that provides a broad outlook on the prevalence of microsporidiosis in Iran. It is necessary to investigate Microsporidia infection in vertebrate and invertebrate hosts and environmental resources in Iran.

Human cryptosporidiosis in Iran: a systematic review and meta-analysis.

Cryptosporidiosis caused by Cryptosporidium spp. is an important parasitic disease that can be life-threatening for children and immunocompromised patients. This systematic review and meta-analysis was designed to determine the prevalence rate of Cryptosporidium infection and related risk factors among the Iranian general population. We searched electronic databases including Google Scholar, PubMed, Science Direct, Scopus and Proquest for articles in English and SID, Magiran, IranMedex, and IranDoc for articles in Persian. Out of 4816 studies identified in the electronic search, 94 articles were eligible for inclusion in the systematic review and meta-analysis. The prevalence rate of cryptosporidiosis by using the random effect model among children, healthy people, and gastroenteritis and immunocompromised patients in Iran was estimated as 3.65, 2.94, 1.29, and 4.54%, respectively. Findings of a phylogenetic analysis inferred by gp60 and 18S ribosomal RNA markers indicated that most of the infection rate belonged to C. parvum (particularly subtype IIaA15G2R1) and C. hominis among understudied groups. The present study is the first systematic review and meta-analysis providing a comprehensive view of the prevalence of human cryptosporidiosis and its related risk factors in Iran. It seems that the awareness of Cryptosporidium prevalence, risk factors, and disease complications may be required for developing effective strategies to prevent infection.

BCL-1 Gene Rearrangements in Iranian Non-Hodgkin Lymphoma Patients.

In the present study, our aim was to assess the incidence of BCL-1 gene rearrangements in formalin-fixed paraffin embedded (FFPE) tissue in patients with non-Hodgkin lymphomas (NHL). The BIOMED-2 protocol was applied to assess the BCL-1 gene rearrangements in NHL patients. PCR amplification was carried out on FFPE in 100 patients with B-cell lymphoma including 89 cases with diffused large B-cell lymphoma (DLBCL) (15 cases under 18 years old) and 11 cases with mantle cell lymphoma (MCL). Out of the 100 patients, 19 cases (19%) were identified to have concurrent translocation involving BCL-1. The significant association was seen between BCL-1 gene rearrangements and the lymphomas in patients older than 55 years (P<0.05). Out of 100 cases, 80 cases were positive and 20 cases were negative regarding CD20. No significant association was found between DLBCL lymphoma in patients under 18 years old and BCL-1 gene rearrangements (P>0.05). In addition, the positive and negative expressions of LCA/CD45 marker were 76% (76/100) and 26% (26/100), respectively. Our findings revealed that BCL-1 gene rearrangement assays using BIOMED-2 protocol can be considered as a valuable approach in detection of the lymphomas.

Genetic Variations of Tumor Necrosis Factor -α-308 and Lymphtoxin-α+252 in Non-Hodgkin Lymphoma and Acute Lymphoblastic Leukemia Patients.

OBJECTIVE(S): Non- Hodgkin lymphoma (NHL) and acute lymphoblastic leukemia (ALL) are two main hematological malignances which have been driven from lymphoid tissue. Genetic polymorphisms in tumor necrosis factor-α (TNF-α) -308 and lymphotoxin-α (LT-α) +252 may affect their transcription and expression which leads to their high plasma level. The frequency of the TNF-α (-308) and LT-α (+ 252) polymorphisms are different for NHL and ALL cases in various populations with different ethnicity. This research is designed to investigate the prevalence and association of TNF-α (-308) and LT-α (+ 252) polymorphisms from NHL and ALL in Azarian patients and healthy individuals from Northwestern part of Iran. MATERIALS AND METHODS: Seventy subjects with ALL and 68 NHL, along with another 130 healthy subjects as control group took part in this study. Genomic DNA was extracted, then genetic polymorphisms in TNF-α and LT-α genes were analyzed with the PCR-RFLP and NCOI as restriction enzyme. A statistical analysis was performed by chi-square test using SPSS software. A P-value of <0.05 was considered statistically significant. RESULTS: A statistically significant difference of LT-α polymorphism was in NHL patients and control (P-value= 0.008) but there was not any association of TNF-α polymorphism between NHL patients and control group. A significant association for TNF-a variant was in ALL and control (P-value =0.005), however, there was no relationship about LT variant between ALL and control. CONCLUSION: The results show that there are significant differences between TNF-α (-308) and LT-α (+252) genetic polymorphisms respectively in ALL and NHL patients with control group from Northwestern part of Iran.

Eosinophilic presentation of acute lymphoblastic leukemia.

PATIENT: Male, 5 Primary Diagnosis: Rule-out appendicitis Co-existing Diseases: Acute lymphoblastic leukemia (ALL) Medication: Chemiotherapy Clinical Procedure: Chest CT • flow cytometry Specialty: Pediatrics' oncology • infection diseases. OBJECTIVE: Rare disease. BACKGROUND: Leukemias are among the most common childhood malignancies. Acute lymphoblastic leukemia (ALL) accounts for 77% of all leukemias. In rare cases, ALL patients may present with eosinophilia. CASE REPORT: Here, a 5-year old boy was admitted to our hospital with a possible diagnosis of appendicitis. This patient's complete blood cell count demonstrated leukocytosis with severe eosinophilia. Following a 1-month clinical investigation, 2 bone marrow aspirations, and flow cytometry analysis, a diagnosis of acute lymphoblastic leukemia was proposed. Finally, the patient was transferred to the oncology ward to receive standard therapeutic protocol, which resulted in disease remission. After chemotherapy for 2 years, patient is successfully treated. CONCLUSIONS: ALL is diagnosed by eosinophilia in rare cases. These patients need immediate diagnosis and intensive therapy due to worsened prognosis of ALL presenting as hypereosinophilia.

Determination of deferiprone in urine and serum using a terbium-sensitized luminescence method.

Optimized conditions, validation and practical applications of a new, rapid and specific fluorometric method for the determination of deferiprone (DFP) in urine and serum samples are reported. The proposed method, which is based on the formation of a luminescent complex with Tb(3+) ion, is evaluated in terms of linearity, accuracy, precision, stability, recovery and limits of detection (LOD) and quantification (LOQ). Under optimum conditions (pH 7.5, [Tb(3+)] = 3 × 10(-4) mol/L, temperature 0 °C and excitation wavelength 295 nm), the relative intensities at 545 nm are linear, with the concentration of DFP in the range 0.072-13 mmol/L for urine and serum samples. The LOD and LOQ, respectively, are calculated to be 0.014 and 0.045 mmol/L for urine and 0.022 and 0.072 mmol/L for serum samples. The intra-day and inter-day values for the precision and accuracy of the proposed method are all < 5%, and the recovery of the method is in the range 97.1-103.8%. The method was applied to human urine and serum samples collected from patients receiving DFP. The results indicated that the method can be successfully applied to the determination of DFP in human urine and serum samples collected for clinical or biopharmaceutical investigations in which simple, rapid, cheap and specific determination methods facilitate and speed up the analytical procedure.