Computational survey of FHIT, a putative human tumor suppressor, truncates structure

Fragile Histidine Triad protein [FHIT], as a known tumor suppressor protein, has been proposed to play crucial role in inhibiting p53 degradation by MDM2. Studies have confirmed FHIT interaction with p53 or MDM2, although functional interacting domains of FHIT with MDM2 and/or p53 are not completely defined. Thus, through determining the significant structural interacting domains of FHIT, information with regard to MDM2 and p53 would be provided. As there were no previous studies evaluating the interaction of optimized important parts of target molecules, docking study was employed. Truncated structures of FHIT were screened to reveal critical sections engaging in FHIT interaction. HEX program was used in order to study the interaction of target structures. Given the total energy, FHIT structures [beta5-7, alpha1] and [alpha1] of FHIT were showed to be better candidates in comparison with other structures in interaction with optimized MDM2 part. Furthermore, FHIT structures [beta4-7, alpha1] and [beta5-7, alpha1] were considered to be better than other structures in interaction with optimized p53 part. FHIT truncates which interact with MDM2 optimized part exhibited lower energy levels than FHIT truncates which interact with p53 optimized part. Our results can be useful for designing new inhibitors of this protein complex interaction which would result in tumor repression

Correlation between cigarette smoking and urine cotinine level in gastric cancer patients

Various substances in cigarette smoke including nicotine have been shown to promote/induce cancer cell proliferation. Since cotinine has a longer half life and stability in the blood, it has become the preferred biomarker for cigarette smoking exposure. Seventy-three gastric cancer patients were included in this study. The tumor tissues were stained with H and E for pathological evaluation. The cotinine levels were measured in urine using a competitive ELISA. Tumors were 90% adenocarcinoma with 63% intestinal and 37% diffuse subtypes. Tumors were poorly [45.2%] or moderately differentiated [41.1%] and localized mainly [77%] in the upper part of stomach. The levels of cotinine were significantly different between smoker [283.83 +/- 178.10 ng/mL] and non-smoker [39.28 +/- 113.34 ng/mL] groups [p < 0.001]. However, there is no-significant correlation between tumor characteristics and cotinine level in smoker patients. Cotinine level correlates with smoking in gastric patients, however, correlation with the tumor features has not been observed

Impact of UGT1A9 polymorphism on mycophenolic acid pharmacokinetic parameters in stable renal transplant patients

There are wide individual differences in pharmacokinetic parameters of mycophenolate mofetil [MMF] among transplanted patients. Some studies have shown that single nucleotide polymorphisms [SNPs] of the Uridine Diphosphate Glucuronosyl Transferase1A9 [UGT1A9] are responsible for these differences in early days after transplantation. Therefore it was decided to evaluate the influence of UGT polymorphism on MMF pharmacokinetics among stable Iranian transplant patients. This was a cross sectional study from March 2008 through December 2008 in Imam Khomeini Hospital affiliated to the Tehran University of Medical Sciences in Iran. Blood samples were taken from 40 de novo stable Iranian renal transplant patients taking 2 g MMF daily with Sr[Cr]

P53 but not cyclin E acts in a negative regulatory loop to control HER-2 expression in MCF-7 breast carcinoma cell line

Cyclin E, HER-2 and p53, are considered as major prognostic markers in breast cancer. As they are related in patho-clinical level, we aimed to check if they have any direct interaction on expression of each other. To study the effect of cyclin E on HER-2 expression, cell lines stably overexpressing cyclin E or its low molecular weight [LMW] isoforms were generated. To understand the results of p53 silencing either alone or in combination with cyclin E overexpression, we created three different p53 stably knocked down cell lines. Protein expression was analyzed by western blot, HER-2 expression in the established cell lines were determined using SYBR green real time PCR and data analyzed by REST software. Results indicate that HER-2 expression is only downregulated following p53 silencing and none of cyclin E isoforms can alter its expression. The presence of cyclin E isoforms in p53 silenced clones also does not altered HER-2 expression. Given the fact that p53 degradation is increased by HER-2 overexpression, these data can draw a regulatory loop in which a non-mutated functional p53 and HER-2 can bidirectionally regulate the expression of these two genes. This study improves our understandings of these pathways and these proteins can be introduced either as a marker or as a target in cancer treatment.

time course of JNK and P38 activation in cerebellar granule neurons following glucose deprivation and BDNF treatment

Low glucose condition induces neuronal cell-death via intracellular mechanisms including mitogen-activated protein kinases [MAPK] signaling pathways. It has been shown that low glucose medium decreases neuronal survival in cerebellar granule neurons [CGNs]. In this study, we have examined the activation of JNK, p38kinase and ERK1/2 pathways in low glucose medium in CGNs. The CGNs were prepared from new-born [P-2 and P-5] rats and cultured in Dulbecco's Modified Eagle's Medium high [DMEM-HIGH] glucose supplemented with Fetal Bovine Serum [FBS] 10% for 7 days. The glucose deprivation was induced through replacing the culture medium with the low glucose [5 mM] medium. The MAPK pathways activation was evaluated through phospho specific antibodies using western blot. The viability of cells was measuring using MTT assay. The results indicated that low glucose reduces the cell survival and brain-derived neurotrophic factor [BDNF] elevates the cell viability in CGNs. The basal c-Jun N-terminal kinase [JNK] activity was high in CGNs and glucose deprivation for 24 h had increased phospho-JNK level to 2-fold compared to basal. BDNF treatment reduced the basal JNK activity within 30 min but had no effect in longer incubations. BDNF also blocked the low glucose-induced JNK activation. In addition, CGNs exhibited high p38 phosphorylation in low glucose medium in 48 h. These results demonstrated that in sustained low glucose conditions, CGNs had high activity of stress-activated MAPK which could induce cellular damage. Moreover, BDNF can prevent JNK and p38 activation in stress conditions and increase cell viability. Our results suggest that in sustained stress conditions, inhibition of JNK and/or p38 pathways might protect neurons from damage in low glucose conditions

Evaluation of the effects of interferon on stability of control genes in huh-7 and HEPG[2] cells

Understanding gene expression variations by using RNA transcript analysis methods during hepatic viral protein interactions with the IFN pathway in hepatic cell lines has recently gained importance. One of the most powerful techniques in gene expression quantification is quantitative real-time RT-PCR. Reference genes used as normalizer in this method may be affected across various experimental conditions or treatments. Hence, in the present study, the influence of IFN-treatment on the mRNA levels of common reference genes including ACTB, GAPDH, TBP, HPRT1 and HMBS was evaluated in Huh-7 or HepG[2] cell lines. Cells were treated with different concentrations of IFN- Then, using geNorm and NormFinder programs, we evaluated the expression stabilities of the above prominent reference genes in three sample groups that included each hepatic cell line and the total data sets. HPRT-1 and GAPDH were the most stable reference genes in the Huh-7 cell line, whereas ATCB, HMBS and GAPDH were the most stable in the HepG2 cell line. TBP was one of the least stable reference genes in the three studied groups. This investigation will provide appropriate reference genes for standardization of quantitative real-time PCR data in an IFN-?stimulated model of hepatocyte cell lines

Enchondroma of the hand: the role of biopsy in the course of diagnosis and treatment

Enchondroma is the most frequent bone tumor of the hand, but chondrosarcoma is rare at this location. There is a high possibility of correct diagnosis of enchondroma and differentiating from its malignant counterpart by precise clinical and radiologic assessment without biopsy, a subject of debate in the literature. At the present study we substantially investigate this problem, in our patients. Case records, radiographs, and histology of 52 solitary enchondroma patients who underwent operation in our hospital between 1998 and 2010, were reviewed. Special attention paid to pre and post -op diagnoses, and compared with each other. Eighty-six percent of our patients were between the second to fourth decades of life, with a slight female predominance. In all, the primary diagnosis of enchondroma according to clinical presentation and radi-ographic appearance, supported by intraoperative gross appearance of tumor, and confirmed histologically by permanent section analysis. There was no mismatch between radiologic and histologic diagnosis. we concluded that correct diagnosis of enchondroma is almost always possible by precise clinical and radiographic assessment with no need for histologic confirmation before definitive treatment

Effects of continuous and interrupted forces on gene transcription in periodontal ligament cells in vitro

The biological mechanisms of tooth movement are based on the response of periodontal tissues to mechanical forces. The final result of these responses is remodeling of the extracellular matrix. Tissue reactions may vary depending upon the type, magnitude and duration of the applied forces. The purpose of the present study was to analyze the effects of centrifugal force on the transcription of collagen type-I [Col-I], matrix metalloproteinase-1 [MMP-1], and tissue inhibitor of metalloproteinase-1 [TIMP-1] genes in human periodontal ligament [PDL] fibroblasts. Human fibroblasts obtained from the PDL were cultured and subjected to centrifugal forces [36.3 g/cm2] for 30, 60 and 90 min continuously. This was also carried out interruptedly, three times for 30 min and six times for 15 min. The mRNAs encoding for Col-I, MMP-1, and TIMP-1 were quantified using RT-PCR. The mRNA levels of Col-I and MMP-1 were increased when continuous force was applied for 30 min and 60 min respectively. The interrupted force had almost no effect on Col-I, MMP-1 and TIMP-1 genes. These results indicate that continuous forces may have a greater effect in inducing gene expression during the remodeling process of PDL compared to interrupted forces with short rest periods

Evaluation of potency of measles vaccine used in Iran: comparison of WHO and NIBSC method in cell culture

Measles has been a major cause of illness and death in children and vaccination against the disease is part of the WHO global immunization program. A suitable vaccine should create maximum immune response against the pathogen and must be safe for the user. Thus, after production, vaccines must be analyzed and controlled by the producer and confirm by relevant governmental organizations. The Food and Drug Control Lab [FDCL], Ministry of Health, is the secondary control center on potency of vaccines in Iran. In this study, we have set up the WHO and NIBSC methods in FDCL and compare these methods on determining the potency of measles vaccine. Measles vaccines were obtained from Razi Institute Iran. Nine dilutions of vaccine [10[-1] to 10[-5]] in 0.5 log interval were mixed with Vero cell suspension and seeded. In WHO method, the cells were incubated at 36°C for 10 days, during which the cells were checked for cytopatic changes everyday. To set up the assay, we tested the vaccine dilution with four different cell suspensions [2x10[5]-5x10[4]/well] and four different concentration of serum [2.5-10%]. Based on our results, in the assays, 5% serum and 1x10[5] cells were used. The potency assay was performed with six different vaccines produced in one batch and the mean potency for Measles was 10[4.32 +/- 0.24] CCID[50]/vial for a ten-dose vial. In NIBSC method following seeding of Vero cells, the medium was removed after 3 hours and overlay was added. Then the plates were incubated at 35°C for 10 days. After incubation period, the overlay was removed, the plaques were stained with methyl violet and counted. This assay was repeated three times and the mean of the results was 5.83 +/- 0.03 log[10] PFU/dose. In this study, we have set up the WHO and NIBSC methods and results indicated that the potency of the vaccine is in acceptable range in either method. Furthermore, the WHO method is simple and less time consuming compared to NIBSC which is complicated and requires more effort to produce reproducible results