Effect of the receptor activator of nuclear factor [ka]B and rank ligand on in vitro differentiation of cord blood CD133[+] hematopoietic stem cells to osteoclasts

Objective: receptor activator of nuclear factor-kappa B ligand [RANKL] appears to be an osteoclast-activating factor, bearing an important role in the pathogenesis of multiple myeloma. Some studies demonstrated that U-266 myeloma cell line and primary myeloma cells expressed RANK and RANKL. It had been reported that the expression of myeloid and monocytoid markers was increased by co-culturing myeloma cells with hematopoietic stem cells [HSCs]. This study also attempted to show the molecular mechanism of RANK and RANKL on differentiation capability of human cord blood HSC to osteoclast, as well as expression of calcitonin receptor [CTR] on cord blood HSC surface

Materials and Methods: in this experimental study, CD133[+] hematopoietic stem cells were isolated from umbilical cord blood and cultured in the presence of macrophage colony-stimulating factor [M-CSF] and RANKL. Osteoclast differentiation was characterized by using tartrate-resistant acid phosphatase [TRAP] staining, giemsa staining, immunophenotyping, and reverse transcription-polymerase chain reaction [RT-PCR] assay for specific genes

Results: hematopoietic stem cells expressed RANK before and after differentiation into osteoclast. Compared to control group, flow cytometric results showed an increased expression of RANK after differentiation. Expression of CTR mRNA showed TRAP reaction was positive in some differentiated cells, including osteoclast cells

Conclusion: presence of RANKL and M-CSF in bone marrow could induce HSCs differentiation into osteoclast

Investigation of leukemia frequency in children of Qazvin province and its correlation with gender, age, and blood groups between 2006-2016

Background: About 8 percent of all cancers in human population are related to leukemia and it is one of the most common malignancies in children. The aim of this study was to compare the prevalence of age, gender and blood group types with the frequency of leukemia among the children with leukemia in Qazvin province during the 2006 to 2016

Materials and Methods: This was a cross-sectional analysis. Investigated population was 110 children and adolescents under 18 years in the hospitals of Qazvin province. The date collecting method was through review of medical records of the patients and their analysis performed by using SPSS version 16

Results: According to data from this study, leukemia ALL-L1 is more frequent in Qazvin than other types of leukemia, and children with ages 0-5 years was more than other age groups. This disorder is more common in boys than girls, and among the patients, the people who has A and O blood groups, and Rh + are the most abundant

Conclusion: such factors like age, gender and blood groups can use as prognostic factors in children leukemia. So that leukemia in children less than 5 years old is more than any other age. In addition to that; the incidence of leukemia ALL-L1 reduced with increasing age in the general population in Qazvin and number of boys with leukemia is more than girls

Ineffectiveness of methylation in rgulation of VHL, ECAD, and RUNX3 genes in erythroid cells differentiated by erythropoietin

Vast variety of intermediate factors including cell cycle regulators, growth factors, transcription factors, and signaling pathways are involved in hematopoietic stem cell [HSC] commitment and differentiation into distinct lineages. VHL, Ecad, and RUNX3 are among these. Epigenetics is currently introduced as a potential mechanism to control the gene regulation. The aim of this study is to reveal the correlation between the expression level and methylation pattern of mentioned genes after in vitro differentiation of cord blood HSCs into erythroid lineage mediated by erythropoietin. After isolation and expansion, the CD34+ cord blood stem cells were divided into two parts. The first part was used to extract the DNA and RNA and the second to differentiate into erythroid lineage. Methylation specific PCR [MSP] and Real-time PCR were used to determine the methylation status and expression levels of the genes, respectively. Although the significant upregulation observed for VHL and Ecad genes and a down-regulation for RUNX3 gene after differentiation, no remarkable changes were seen in methylation pattern compared with cord blood HSCs by MSP technique. It is appearing that methylation pattern in promoter region has not an effective role in expression of VHL, Ecad, and RUNX3. Moreover, considering the inability of MSP method to detect subtle differences in methylation level a more sensitive method is needed to distinguish the methylation levels of these genes before and after erythroid differentiation

Lentiviral vectors titration using real-time PCR

Lentiviral vectors [LVs] are useful vehicle for genetransfer to dividing and non-dividing cells and genetic manipulations. However, the use of lentiviruses in studies requires an accurate titration technique.Quantitative real-time PCR [qPCR] is a sensitive technique for the indication and quantitation of retrovirals particles. In this study, we used the qPCR for lentiviral vector titeration. The puromycin resistance gene as templates for an SYBR green-based real-time qPCR method and detect lentiviral copy number integrated lentiviral DNA. Consequently, this studyshowed that theusing ofantibioticresistance genesviral particles titration maybeefficient with highly accuracy

Survey of clinical features, pathogenesis and therapeutic options for Ebola haemorrhagic fever

The genus Ebola virus first was recognized in 1976, when two outbreaks occurred in Zaire and Sudan. Ebola virus disease [EVD] is a highly contagious disease that can affect both human and nonhuman primates: Zaire ebolavirus [ZEBOV], Sudan ebolavirus [SEBOV], Côte d'Ivoire ebolavirus [CEBOV], Bundibugyo ebolavirus [BEBOV] and Reston ebolavirus [REBOV] are five members of the Filoviridae family that can cause haemorrhagic fever. EVD is transmitted by direct contact with contaminated blood or other biological fluids of the infected animals such as chimpanzees, gorillas, fruit bats, monkeys, forest antelope and porcupines found ill or dead or in the rainforest. Ebola is responsible for different clinical futures that can be ranged from fever, headache, arthralgia, myalgia, abdominal pain, anorexia and vomiting to severe respiratory disorders, viral hemorrhagic fever, cardio-vascular disorders and hypovolaemic shock. Although there is no specific treatment for EVD, considerable advances like use of monoclonal antibody, intefron and Favipiravir/T-705 as effective chemotherapeutic agent in treatment of EBV have been made. To date, 25 outbreaks of EVD have been reported. Hence, EVD as a zoonotic disease should be more focused not only in endemic area but also in throughout the world. Awareness of the disease and routes of transmission and also continuous surveillance to combat disease and outbreaks is necessary

Correlation between methylation and expression level of P15 and P16 genes during differentiation of cord blood stem cells into erythroid lineage mediated by erythropoietin

Several influential factors such as transcription factors and intracellular signaling components are involved in differentiation of stem cells into a specific lineage. P15 and p16 proteins are among these factors. Accumulating evidences has introduced the epigenetic as a master regulator of these factors during lineage specification. The main objective of this study is to determine the correlation between the expression level and methylation pattern of P15 and P16 genes in erythroid lineage after in vitro differentiation by erythropoietin [EPO]. The purified and expanded CD34+ cord blood stem cells were differentiated into erythroid lineage in the presence of EPO. DNA was isolated from both cord blood stem cells and differentiated cells. The Real-Time PCR performed using cDNA and the isolated DNA was used in methylation Specific PCR [MSP] reaction for methylation pattern analysis in both pre and post differentiation stages. The study demonstrated that P15 and P16 genes have partial methylation after erythroid differentiation by EPO. The Expression of P15 gene was higher after differentiation and the expression of P16 gene had a slightly decreased level in post differentiation stage. Significant increase in P15 gene expression after differentiation to erythroid lineage, suggests the remarkable efficacy of this gene in erythroid function. According to upregulation of P15 gene after differentiation despite unchanged methylation status and slight down regulation of P16 gene with slight hyper-methylation of the gene it can be suggested that although the methylation can affects the expression level of P16 gene, the P15 gene is not affected by this mechanism during erythroid differentiation mediated by EPO

Profile of side Effects in Patients Receiving Blood Transfusion from the Perspective of Management Unit

Background: In spite of being vital to save the patients' life, blood transition may be dangerous and even fatal, too the aim of this study was to investigate the side effects [complications] of blood transfusion in the educational hospitals of Qazvin.

Materials and Methods: This is a descriptive cross sectional and practical study that was carried out in 2010. In this study, all the patients of four health training centers in Qazvin, that have had blood transfusion and complications, were considered as a part of the statistical community. The instrument for data collection was checklist which was filled through an interview with blood bank manager and some other responsible individuals and scrutinizing files of patients who had blood injection among the blood products consumption, request for the packed cells was the most and for fresh frozen plasma was the least.

Results: 75% of these people had only one blood injection and the maximum injection volume was 100cc which was done mostly in the evening. Most of the transfusion history belonged to 21-30 year olds in our statistical community. 56% of all Patients that had transfusion, possessed background of some disease such as heart problems [21.9%]. More than half of them [2.56%] had a chill feeling complication transfusion and there was a significant relationship between the blood transfusion volume and itching complication.

Conclusion: Existence of a continuous association between blood transfusion organization and hospitals is indispensable. Therefore, it seems that Hemovigilance system or computer connected network to send reports, between hospital centers and blood transfusion organization of Iran, can be an appropriate solution.

Prevalence of blaCTX-M gene in multi-resistant escherichia coli isolated from urinary tract infections, Tehran, Iran

The emergence and increase in the incidence of Extended-spectrum beta lactamase [ESBL] producing Escherichia coli [E. coli] has become an emerging challenge especially in hospitalized patients with urinary tract infection [UTI]. The aim of the present study was to survey the frequency of bla CTX-M genotype in ESBL producing E. coli isolated from hospitalized patients with urinary tract infection and determination of their antibiotic resistance pattern. A total of 135 E. coli isolates were collected and isolated from patients with UTI. The isolates were subjected to confirmatory phenotype tests for the presence of ESBL. 75 E. coli isolates were confirmed as ESBL-positive by double disc synergy test. In vitro susceptibility of ESBL isolates to 15 antimicrobial agents amoxicillin, penicillin, ceftazidime, cefotaxime, cefoxitin, ceftriaxone, cefixime, cephalexin, co-trimoxazole, gentamicin, nalidixic acid, ciprofloxacin, nitrofurantoin, amikacin, and imipenem was performed by Kirby-Bauer's Disk diffusion method according to Clinical and Laboratory Standards Institute [CLSI, 2012] guideline. PCR method was used to identify bla CTX-M gene in 75 ESBL positive strains. PCR and sequence analysis showed that 75 [55.5%] isolates produced bla CTX-M genes. In vitro susceptibility of ESBL producing E. coli showed that all of them were resistant to amoxicillin and penicillin. The rates of resistance to the majority of tested antibiotics varied among 61% to 100%, with the exception of amikacin [14.7%] and imipenem [2.7%]. Our results showed that the frequency of bla CTX-M was strikingly high [93.3%] in patients with UTI. These data confirmed that the frequency of bla CTX-M genes was high among E. coli isolated from patients with UTI. The trend of multidrug-resistant profile has been associated with bla CTX-M gene is alarming. Therefore, it is very important to establish a routine screening of ESBL in clinical isolates to prevent dissemination of resistant isolates in health care settings

Characterization of coagulase-negative staphylococci isolated from hospitalized patients in Tehran, Iran

Coagulase-negative staphylococci [CNS] are a main cause of nosocomial infection. The main purpose of this study was to determination of frequency of CNS isolates in in hospitalized patients and their susceptibility pattern to antimicrobial agents. During 11 month study, 65 CNS clinical isolates were recovered from hospitalized patients in different wards of hospital. In vitro susceptibility of isolates to 12 antimicrobial agents Penicillin; Ampicilin; Cephalothin; Cefoxitin; linezolid; Nitofurantoin; Erythromycin; Norfloxacin; Gentamicin; Vancomycin; Chloramphenicol and Oxacillin was performed by Kirby-Bauer's Disk diffusion method according to Clinical and Laboratory Standards Institute [CLSI] criteria. Out of 1875 samples of hospitalized patients 65[3.47%] patients were infected with CNS. Twenty one [32.3 %] were isolated from the urine samples, 17[26.1%] from sputum, 15[23.1%] from pus samples, 8[12.3 %] from ear swabs, 3[4.7%] from fluid and 1[1.5%] from blood sample. All of CNS isolates were sensitive to nitrofurantoin. The rates of resistance to the majority of antibiotics tested varied between 4.5% and 100 %. The rate of resistance to beta lactam antibiotics, Chloramphenicol, erythromycin, gentamycin was high [more than 70%]. The most of isolates remained susceptible to linezolid, and nitofurantoin. All of isolates were susceptible to vancomycin. Multi-drug resistant CNS with reduced susceptibility to linezolid and nitrofurantoin are emerging pathogens of clinical concern. Monitoring of antibiotic resistance with attention to multi-resistant profile and aware to practitioners in the field is necessary

Study of flagellin profiling in multidrug resistant Pseudomonas aeruginosa [MDRPA] isolated from burn wound infections, Tehran, Iran

Nosocomial infections of multidrug-resistant Pseudomonas aeruginosa [MDRPA] are a growing concern in hospitalized patients in burn centers. The aim of this study was to investigate the flagellin profiling and antibiotic susceptibility of P. aeruginosa isolated from burn wound infections. During 8 month study, 73 clinically P. aeruginosa isolates collected from patients hospitalized in burn ward. P. aeruginosa isolates were identified using standard laboratory procedures. In vitro susceptibility of clinical isolates of P. aeruginosa to 6 antimicrobial agents were investigated by Clinical and Laboratory Standards Institute [CLSI 2012] Kirby-Bauer disk diffusion assay. The frequency of different type of flagellin was investigated by using specific primers and by PCR method. The resistance rates of our isolates to 6 tested antimicrobial agents were relatively high. Imipenem has good activity while tobramycin and ciprofloxacin do not have any effect on P. aeruginosa isolates. Of 73 isolates 59 [80.8%] were multidrug resistant. Twenty eight of 73 isolates were resistant to all antibiotics. Agarose gel electrophoresis of chromosomal DNA exhibited that 59 isolates [80.8%] of P. aeruginosa had type A flagellin while only 14 isolates [19.2%] had type b flagellin. Given the antibiotic failure treatment, it appears that alternative ways such as immunity to prevent of these infections could be informative. Our survey of flagellin profiling of multidrug-resistant P. aeruginosa isolates exhibited high frequency of type a flagellin as a major virulence factor has important role of immunity against infections caused by MDRPA. Functional surveillance of multidrug-resistant P. aeruginosa in order to prevention of resistance dissemination is necessary

Challenges in the treatment of Iranian patients with leukemia in comparison with developed countries from the perspective of specialists

Evaluation of the factors associated with treatment process of leukemia and comparison with current related approaches in developed countries can present a good indicator to assess the weak and strong points in healthcare system of our country in leukemia treatment. The objective of this research is general and specific description of the challenges and shortcomings in Iranian healthcare system and monitoring of hematologic malignancies as well as comparison with developed countries. Our study is a descriptive-cross-sectional study. 100 hemato-oncologist , pathologists, and faculty members throughout the country were selected by random cluster sampling. Data collected using questionnaires with Cronbach's alpha coefficient of 0.76 . SPSS and Chi-square test were used for data analysis. According to the specialists, lack of advanced diagnostic facilities as well as cell and BM banks together with high treatment expenses are the main factors contributing to poor treatment processes in Iran, which are far from worldwide standards. The use of novel currently methods used in developed countries for leukemia treatment, financial and psychological support of patients under treatment , making underprivileged provinces well-equipped ,balanced specialist service distribution relative to capital city either in diagnosis or treatment are factors which makes system standardized. Moreover, integrated institutional work in relation to leukemia incidence and statistical analysis of mortality and morbidity rate can pave the way for reducing and eliminating the problems in diagnosis and treatment of leukemia patients