Association of the co-stimulatory molecules polymorphisms with CMV infection in liver transplant recipients

Co-stimulatory molecules play a critical role in regulating T-cell function during CMV infection after liver transplantation. To investigate the relationship between the polymorphisms of the co-stimulatory genes and the susceptibility to CMV infection after liver transplantation. Single nucleotide polymorphisms [SNPs] in PD-1 gene [PD1.1 A/G, PD1.3 A/G, PD1.9 C/T] ICOS [-693 A/G, 1720 C/T], CTLA-4 gene [-318 C/T, 1722 T/C, 1661 A/G, 49 A/G] and CD28 [+17 C/T] were analyzed by PCR-RFLP in 70 liver transplant patients. CMV infection was determined in these patients by antigenemia test. CTLA-4 49G showed significant association with CMV infection [p=0.03, OR=3.82, 95% CI: 0-3.5; p=0.01, OR=004, 95% CI: 0-1.3]. G and T alleles in CTLA-4 gene [-318 C/T and 1661 A/G] [p=0.03, OR=0, 95% CI: 0-3.5; p=0.01, OR=0.04, 95% CI: 0-1.3] were significantly higher in CMV-infected rejector group. CTLA-4 have significant role in CMV pathogenesis and rejection among CMV-positive liver transplant patients

molecular and antigenic tissue impact of viral infections on liver transplant patients with neonatal hepatitis

Pathogenesis of neonatal hepatitis relates to various underlying causes including viral infections. Both hepatotropic and non-hepatotropic viruses may induce liver failures in infants before birth, during delivery, or shortly after birth. The tissue impact of HCMV, HSV, HBV, HCV, and rotavirus and adenovirus infections was evaluated in studied infants with neonatal hepatitis. The history of viral infections was analyzed in paraffin-embedded biopsy and autopsy tissues of 22 infants with neonatal hepatitis between years 1996 and 2007, retrospectively. The tissue molecular presentation of HBV, HCV, HCMV, HSV, adenovirus, and rotavirus was evaluated by different qualitative simple and nested PCR and RT-PCR protocols. Immunohistochemistry [IHC] method was used for studying the antigenic prevalence of HSV-1, 2; HBV, HCMV and adenovirus infections. Also the laboratory liver indices of all patients with neonatal hepatitis were analyzed. The HBV and HSV genomes were detected in 3 [14%] of 22 infants. The rotavirus and HCV-RNA and also the HCMV-DNA were detected separately in 1 [4%] of 26 paraffin-embedded autopsy and biopsy tissues. The HBV and HSV-1 specific antigens were separately diagnosed in 1 [4%] of 26 neonatal samples by IHC protocols. Also the HSV-2 antigen was seen in 5 [23%] of 22 liver autopsy and biopsy specimens. Co-infections with HCMV, HSV, HBV, HCV, and rotavirus were detected in these infants with hepatitis. Diagnosis of single and mixed molecular and antigenic traces of HCMV, HSV, HBV, HCV and rotavirus underlines the etiologic role of these viruses in clinical pathogenesis of neonatal hepatitis

[Cloning and expression of vascular endothelial growth factor receptor-1 in Escherichia coli and analysis of its growth inhibition effect on human umbilical vein endothelial cells]

Angiogenesis a process that results in neo-vascularization is an essential stage in growth of solid tumors and the formation of metastases. Vascular endothelial growth factor [VEGF] and its receptors, VEGFR1 [Flt-1] and VEGFR2 [KDR/Flk-1], are the major regulators for tumor angiogenesis. Recent studies showed that second domain of VEGFR-1is a key factor for VEGF/VEGFR-1 interaction. In this study, after RNA purification and cDNA synthesis, the second domain of VEGFR-1 [VEGFR-1-II] was amplified by PCR and cloned in T/A cloning vector. In order to increase the expression of the protein, we sub-cloned the gene into pET22b[+] and transformed the construct in Rosseta-gami 2, an efficient host for expression. The expression was induced by IPTG and confirmed by SDS-PAGE and Western Blotting. The recombinant protein was purified by IMAC column and the growth inhibition of human umbilical vein endothelial cells [HUVEC] was analyzed by the recombinant protein. The results of SDS-PAGE and Blotting confirmed the protein purification accuracy. The recombinant protein concentration was determined by Bradford protocol. The results showed that nearly 300 micri g/L VEGFR-1-II protein was produced. The function of this protein was confirmed by inhibition of HUVEC cells growth. Since this protein inhibited the angiogenesis in vitro it may be consider as an efficient anti-angiogenesis factor

Tolerance induction by CD40 silenced dendritic cells through antisense

One of the valuable tools for inhibiting the specific gene expression is antisense technique. To determine T cell responses, co-stimulatory molecule expression on the antigen presenting cells is important. In the present study, the effects of high affinity antisense against CD40 mRNA on the function and phenotype of DCs [dendritic cells] were investigated. The DCs were separated from the mice spleens and then cultured in vitro. By means of lipofectamine 2000, the antisense was delivered into the cells and the efficacy of transfection was estimated by flow cytometry. Also, the mRNA expression and protein synthesis were assessed by real time PCR and flow cytometry, respectively. The DCs were transfected with 6 M antisense and 2 l lipofectamine 2000. The percentage of CD40 expression in DCs was 38%. The results showed that CD40 expression is reduced in DCs to 22% and 24%. By annexine V and propidium iodine staining, we could evaluate the viability of the transfected cells. The inhibition of CD40 gene expression was associated with the increase in IL-4 secretion. This shifted the DCs to stimulate Th2 cytokine production from the allogenic T cells. In addition, in the MLR, the DCs without CD40 expression showed poor allostimulatory effects. This finding is valuable in the study of the costimulatory molecules of DCs. These data demonstrate that direct interference of the cell surface expression of CD40 at transcriptional level by antisense confers tolerogenecity potential of DCs. This approach is a useful tool through which DCs become tolerogenic and can be studied as a potential therapeutic option for the autoimmune diseases and allograft rejection

[ survey on comparison of tympanic and nasopharyngeal temperatures in patient undergoing open heart surgery]

Body temperature controlling in patient undergoing open heart surgery is very important and critical. In fact it is the base of work and by measuring temperature correctly, complications of hypothermia can be prevented. The overall purpose of this study was to determine and compare tympanic and nasopharyngeal temperatures in patient undergoing open heart surgery with hypothermia. This was a correlation study. The sample consisted of 60 patients undergoing open heart surgery with hypothermia. Body temperatures are measured in three sites - right ear, left ear and nasopharynx. Both of them were measured simultaneously before, during and after hypothermia. Tympanic temperatures were measured with an infrared thermometer and the nasopharyngeal temperatures were monitored by heart and lung machine, in fact it is a process in open heart surgery. This study showed that the mean of body temperatures are different in three sites. The difference between right and left ear wasn't significant, but the mean of nasopharyngeal temperature was significantly different with right and left ear. In order to determine the sensitivity of tympanic to changes of temperature, correlations between three sites - right ear, left ear and nasopharyngeal - are calculated. Temperatures at three sites had a high correlation [p= 0.01]. All sites are sensitive to changes of body temperatures and they can show core temperatures of body well. Therefore, when there are limitations for monitoring of body temperatures by nasopharyngeal route, the tympanic route can be a good replacement

Teaching styles in clinical nursing education: a qualitative approach

Background and purpose: There are many studies about nursing clinical settings and their problems, but the teaching style of teachers on the bedside has not been studied as a whole. Therefore, this study was conducted to assess, describe and interpret nursing trainers' perceptions of the teaching style in clinical settings

Methods and Materials: The grounded theory approach was used to conduct this study. Fifteen nursing teachers were interviewed individually in 2006-2007. The interview protocols were tape-recorded and later transcribed verbatim. The transcriptions were analyzed using Strauss and Corbin's method

Results: Three major themes and 12 sub-themes emerged from the study data which portray the clinical teaching styles of the nursing teachers. The main themes are multiple styles in teaching, nature of clinical teaching, control and adaptation in the educational atmosphere and multiplicity in teaching style. Individualized styles were observable across teachers, but they varied across situations, type of skills [content], educational environment and facilities, levels of the learners, and the control and accommodation of teachers with the teaching atmosphere

Conclusion: Although teaching style is a complex phenomenon, but this study has helped emerge some of the rules and principles of clinical training of nurses

[ comparison of the efficacy of intra-articular injection of morphine with marcaine in patients to relief the pain associated with knee arthroscopy]

Knee arthroscopy is an approved technique for the diagnosis and treatment of intra-articular lesions. Moderate to severe pain is experienced after surgery; thus, relieving pain post arthroscopy, will help patients in performing their daily activities as soon as possible. Many studies have been performed for reducing pain after arthroscopy. The aim of this study is to compare the efficacy of intra-articular injection of morphine with marcaine in patients for pain relief after arthroscopy. 30 patients were considered for arthroscopic surgery, due to the tearing of the menisci. In this simple non-probability trial, patients were divided in two groups. The first group received 7cc intra-articular marcaine at 0.5% and the second group received 10mg of intra-articular morphine after the arthroscopy. The response was measured by VAS in hours 6, 12, 18, 24 postoperatively and by flexion, extension and walking. The results showed that there was no significant statistical difference between the two groups, except in hour 6 after surgery, indicating marcaine is more effective than morphine. There were no side effects experienced within the two groups. Age, gender, height and weight also had no effect in reducing the pain in patients. Intra-articular Injection of marcaine is more effective than morphine six hours after surgery; however, there are no differences between them after that time frame. More research is needed in order to reduce pain after arthroscopy