[Metformin reduces cisplatin-mediated apoptosis in gastric adenocarcinoma cells]

Metformin, a drug widely used for type 2 diabetes, may also have anti-cancer properties. The purpose of this study was to examine the effect of metformin on cisplatin cytotoxicity in the gastric adenocarcinoma cells line [MKN45]. In this study, cells viability and apoptosis were measured using the [3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide] assay and flow cytometry, respectively. Moreover, the expressions of mammalian target of rapamycin, survivin and AKT genes were evaluated by RT-PCR. All experiments were performed in triplicate. The results showed that each of metformin and cisplatin separately reduced the viability of cancer cell, but in co-administration, metformin reduced the cytotoxicity of cisplatin. In co-administration, the survivin expression was increased followed by a reduction in cisplatin anti-cancer effect. Therefore, the antagonistic effect of drugs can be associated with survivin expression. The results also revealed that the anti-apoptotic effects of metformin co-administrated with cisplatin are associated with increased AKT expression. It seems that in gastric cancers, metformin is not an appropriate choice to make cells sensitive to cisplatin and the antagonistic effects of the two drugs should be considered when they prescribed in combination

[ effectiveness of group metacognitive therapy on anxiety thought and improvement thought control strategy in patients with generalized anxiety disorder]

Generalized anxiety disorder [or GAD] is defined as an excessive, exaggerated anxiety and uncontrollable worry. The purpose of this study was to investigate the effectiveness of group metacognitive therapy [G-MCT] on Anxiety thought and improvement of thought control strategy in individuals with generalized anxiety disorder [GAD]. The study population consisted of all outpatients with GAD presenting to clinics in Tehran counselling center. A number of the patients conveniently were selected. Then using of Metacognition Questionnaire [MCQ-30], Thought Control Questionnaire [TCQ] and anxiety thought inventory [ANTI] and the Structured Clinical Interview for DSM-IV Disorders [SCID-I], 30 females were selected and randomly were assigned in two groups [15 in experimental group and 15 in control group]. From 15 patients of experimental group, 12 patients completed the treatment. Group metacognitive therapy was administered to the experimental group in ten, 120 minutes sessions during 2.5 months period. All subjects completed questionnaires before and after intervention and in follow-up period after 3 month. The results of ANCOVA test showed that Group metacognitive therapy have significant effect on negative metacognitive beliefs. The results of ANCOVA test also showed that Group metacognitive therapy significantly have decreased anxiety thought, and improved thought control strategy. These results remained in 3 months follow-up period. Group metacognitive therapy has some more benefits on metacognition beliefs and anxiety thought, and improved thought control strategy

Cytogenetic and FMS-like tyrosine kinase 3 mutation analyses in acute promyelocytic leukemia patients

The secondary genetic changes other than the promyelocytic leukemia-retinoic acid receptor [PML-RARA] fusion gene may contribute to the acute promyelocytic leukemogenesis. Chromosomal alterations and mutation of FLT3 [FMS-like tyrosine kinase 3] tyrosine kinase receptor are the frequent genetic alterations in acute myeloid leukemia. However, the prognostic significance of FLT3 mutations in acute promyelocytic leukemia [APL] is not firmly established. In this study, the chromosomal abnormalities were analyzed by bone marrow cytogenetic in 45 APL patients and FLT3 internal tandem duplications [ITD] screening by fragment length analysis and FLT3 D835 mutation by melting curve analysis were screened in 23 APL samples. Cytogenetic study showed 14.3% trisomy 8 and 17.1% chromosomal abnormalities other than t[15;17]. About 13% of the patients had FLT3 ITD, and 26% had D835 point mutation. FLT3 ITD mutation was associated with higher white blood cell count at presentation and poor prognosis. The PML-RARA translocation alone may not be sufficient to induce leukemia. Therefore, we assume that FLT3 mutations and the other genetic and chromosomal alterations may cooperate with PML-RARA in the development of APL disease

Evaluation of 56 cases of long-segment anastomosis of LITA to LAD in Rajaei heart center

Long-segment reconstruction of the diffusely diseased left anterior descending artery [LAD] with left internal thoracic artery [LITA] is one of the methods offered in order to deal with complicated, multiple, and long-segment lesions in the LAD. In this prospective study, we analyzed the results obtained with this technique. Between Feb. 2007 and Feb. 2009, 56 patients underwent surgery via this technique. The LITA was used as a patch along the opened narrow segment of the LAD from 2 to 8 cm. Data on all the patients were collected, and all the patients were worked up for postoperative complications such as postoperative myocardial infarction, ECG changes, NIHA class, enzymatic changes, and postoperative bleeding. CT-Angiography was performed between 6 to 1 8 months after surgery in some cases. Fifty-six cases, comprising 42 [75%] men and 14 [25%] women between 43 and 78 years of age [mean age= 59.8 +/- 9.3 years] with multiple and long-segment lesions in the LAD were included in this study. Preoperative risk factors were hypertension [66.1%], diabetes [57.1%], hyperlipidemia [50%], cigarette smoking [50%], renal failure [1.8%], and positive family history [7.1%]. Twenty-three [41.1%] patients had remote and 9 [16.1%] had recent myocardial infarction. Significant left main lesions were found in 7 [12.5%] patients, peripheral vascular disease in 3 [5.3%], and preoperative arrhythmias in 2 [3.6%]. The mean number of grafts was 2.85 +/- 1.5. Postoperative complications were arrhythmias in 10 [1 7.8%] patients, postoperative myocardial infarction in 1 [1.8%], surgical bleeding in 7 [12.5%], infections in 3 [5.3%], plural effusion in 3 [5.3%], tamponade in 2 [3.6%], and pericardial effusion in 1 [1.8%]; there was no mortality amongst the patients. CT-angiography, performed in 6 patients between the six and eighteenth postoperative months, revealed patent anastomoses in all the patients. Long segment and multiple lesions in the LAD pose a challenge for cardiac surgeons. The results of long-segment LAD reconstruction using the LITA are very encouraging

[Detection of viability and metabolic activity of NB4 cell line treated with AZT]

APL is a Prevalent leukemia that Approximately included 5-10% of patients with acute myeloblastic leukemia. ATRA and recently arsenic is used for treatment. ATRA leads to resistance to treatment and arsenic is toxic in high doses. AZT induce cell death in different ways. The purpose of this study was Assessment of effect of AZT, a telomerase inhibitor, on NB4 cell line [APL cell line] to reduce toxic effect of high dose arsenic. In this study, viability and metabolic activity of NB4 cells, treated by different concentrations of AZT[50,100,200 micro M], was assessed by trypan blue dye method and MTT assay respectively. Treated cells with AZT=50,100,200 micro M showed decreased viability, both in dose-dependent and time-dependent through trypan blue dye method and decreased cell metabolic activity by MTT assay. Considering that AZT is able to induce apoptosis and decrease cell activity, it seems AZT is a suitable drug for inhibiting the growth of tumor cells

[Combination effect of low dose of arsenic trioxide and AZT on viability and metabolic activity of NB4 cell line]

Acute promyelocytic leukemia [APL] is a distinct subtype of acute myeloid leukemia. APL is characterized by a balanced reciprocal translocation between chromosomes 15 and 17, t[5;17]]. Important therapeutic strategies for this disease are ATRA and Arsenic trioxide. To eliminate tumor cells with arsenic, high dose of arsenic is needed. But high dose is toxic for normal tissue. The purpose of this study is Assessment of effect of low dose of arsenic trioxide in combination with AZT on NB4 cell line [APL cell line] to reduce toxic effect of high dose arsenic. In this study after NB4 cell line culture and proliferation, the cells treated with low dose of arsenic trioxide[0.5 micro M] in combination with different doses of AZT[50, 100, 200 micro M] and then viability and metabolic activity was assessed by try pan blue and MTT assay respectively. Low dose of arsenic [0.5 micro m] alone and in combination with dose of 50 micro m of AZT has little effect on viability and metabolic activity but in combination with higher dose of AZT has significant effect on viability and metabolic activity and both viability and metabolic activity significantly reduced. Different apoptosis- induced mechanisms cause apoptosis by arsenic and AZT. Since some of these mechanisms between AZT and arsenic are similar, so maybe these similar mechanisms cause synergic effect and significant reduction of viability and metabolic activity in combination of these two drugs

Coronary artery revascularization in a young adult with solitary coronary aneurysm probably secondary to childhood Kawasaki disease

Surgical revascularization for coronary artery lesions secondary to Kawasaki disease [KD] has been rarely reported in adolescent patients. We report a young adult with no coronary risk factors but with a giant solitary coronary aneurysm with obstructive thrombosis inside, presumably secondary to KD, who underwent coronary artery bypass grafting [CABG] with left internal thoracic artery [LITA] and SVG. Because coronary artery sequelae of KD can be a cause of ischemic heart disease even in young adults, heightened awareness of this entity is required for young adults with coronary lesions but without coronary risk factors

[ effect of arsenic trioxide treatment on mitochondrial apoptotic gene expression in acute promyelocytic leukemia cell line]

Acute promyelocytic leukemia [APL] is one of the most malignant forms of acute leukemia with a fatal course of only weeks which represents 10-15% of AML in adults. Arsenic trioxide as a single agent factor [without chemotherapy] is the treatment of choice for APL patients; it induces cell death through apoptosis but the mechanism by which arsenic targets apoptosis and dramatically affects gene expression remains poorly understood. Since arsenic is used as first line treatment in Iran, it is worth investigating its effect on expression of genes involved in APL. In this descriptive study, to understand the underlying mechanisms of cell death induction by arsenic, we treated NB4 cell line in a dose and time dependent manner. Extracting RNA and synthesis of cDNA, gene expression of apoptotic genes in mitochondrial pathway including caspase3, Mcl-1 and Bcl-2 was analyzed through Real-Time PCR. Our findings showed that As[2]O[3]-induced cell death was paralleled by reduced expression of the antiapoptotic protein Bcl-2 but the expression of Caspase3 and Mcl-1 did not change after arsenic treatment. These results suggest that changes in Bcl-2 gene expression may be one of the mechanisms of action of arsenic in induction of apoptosis, while Caspase3 and Mcl-1 gene expression are not affected by arsenic at the transcriptional level

effect of remifentanil on the hemodynamic changes of parturients in elective cesarean section by general anaesthesia

Background and Purpose: Since narcotics and benzodiazepine cannot be applied for cesarean anesthesia due to the effect of respiratory depression on the infant, parturients generally experience hemodynamic changes in general anesthesia. Remifentanil is a very short-effected narcotic which can be metabolyzad in the infant's body, and is expected not to cause respiratory depression. Therefore, the present study intends to examine the effect of remifentanil on parturients' hemodynamic changes

Methods and Materials: This clinical trial was conducted on 72 full term parturients with ASA physical status I and II, who were randomly assigned into two groups. Before the induction of anesthesia, the first group received 0.75 micro/gkg remifentanil followed by its infusion of 0.10 µg/kg. The second group received bolus and infusion of normal saline. Heart rate, systolic, diastolic and mean arterial blood pressure were measured before the induction of anesthesia, before laryngoscopy, as well as 1, 3, 5, 15 and 30 minutes after intubation. For dat analysis, independent sample t-test, Mann Whitney U, Chi-square and paired sample ttest were used in SPSS

Results: No significant differences were found between the two groups as for heart rate, systolic, diastolic and mean arterial blood pressure in baseline measurement [p>0.05]. After remifentanil administration, mean heart rate before laryngoscopy as well as 1, 3, 5, 15 and 30 minutes after laryngoscopy were significantly lower than the placebo group. However, systolic, diastolic and mean arterial blood pressure were lower in the first group only on occasional minutes

Conclusion: The present study suggests that the mentioned dosage of remifentanil does not have sufficient clinical effects on controlling parturients' hemodynamic responses during general anesthesia for cesarean section

Multiple cardiac malformation and congenital nephrotic syndrome in two sibbling from Iran

Congenital nephrotic syndrome is a rare disease in Iran but co morbidity with cardiac malformation in two consecutive male siblings with pulmonic stenosis, closed VSD, severe PS with small ASD and sever Right ventricular hypertrophy and tricuspid regurgitation is reported

Frequency of BCR-ABL fusion transcripts in Iranian patients with chronic myeloid leukemia

A specific chromosomal abnormality, the Philadelphia chromosome, is present in 90 - 95% of patients with chronic myeloid leukemia. The aberration results from a reciprocal translocation of chromosomes 9 and 22, creating a BCR-ABL fusion gene. There are two major forms of the BCR-ABL fusion gene, involving ABL exon 2, but including different exons of BCR gene. The transcript b2a2 or b3a2 codes for a p210 protein. Other fusion gene leads to the expression of an e1a2 transcript, which codes for a p190 protein. Other less common fusion genes are b3a3 or b2a3 [p203] and e19a2 [p230]. The incidence of one or other rearrangement in chronic myeloid leukemia patients varies in different reports. In general, fusion transcripts are determined individually, a process which is labor- intensive in order to detect all major fusion transcripts. The objective of this study was to set up a multiplex RT-PCR assay for detection and to determine the frequency of different fusion genes in 75 Iranian patients with chronic myeloid leukemia. Peripheral blood samples were analyzed by multiplex RT-PCR from 75 adult Iranian chronic myeloid leukemia patients to detect different types of BCR-ABL transcripts of the t[9;22]. All patients examined were positive for some type of BCR/ABL rearrangement. The majority of the patients [83%] expressed one of the p210BCR-ABL transcripts [b3a2, 62% and b2a2, 20%], while the remaining showed one of the transcripts of b3a3, b2a3, e1a2 or co-expression of b3a2 and b2a2. The rate of co-expression of the b3a2 and b2a2 was 5%. In contrast to other reports, we did not see any co-expression of p210/p190. Co-expression may be due to alternative splicing or to phenotypic variation, with clinical course different from classic chronic myeloid leukemia

[Study in prevalence of Bartonella henselae infection in domestic cats from Tehran]

In present study the zoonotic role of cat in Bartonella henselae transmission have determined. It has done on 100 cats in 2 groups: indoor and outdoor and in 2 age's subgroups. Bartonella henselae was not isolated from blood culture of cats. 23 cats from 100 cats [23%] had antibodies against B. henselae. In this study there were no significant differences statistically in seroprevalence between cats and their owners [p<0.381]. Seroprevalence of cat owners was 18% and in control group [persons who own no cat] was 5%. There were significant differences [p<0.004] between cat owners and control group. Only 6 cats of 50 cats under 6 months old had antibodies to bartonella henselae, and in the other group 17 cats were seropositive and there were significant differences between these two groups [p<0.009] that showed seroprevalence in cats more than 6 months old is higher than the cats under 6 months old. 2 indoor cats from 50 indoor cats and 21 outdoor cats from 50 outdoor cats were seropositive and comparing of these two groups showed significant differences [p<0.0005], which confirmed indoor cats are less frequently infected than outdoor or stray cats

CMV detection and monitoring in bone marrow transplant [BMT] recipients by Real-Time PCR [RQ-PCR]

Cytomegalovirus [CMV] is an important pathogen in patients undergoing bone marrow transplantation. The prevalence of CMV varies from 30-100% in different countries as shown by seroepidemiological studies. Only 20-25% of patients develop CMV disease. Because of the similarity between CMV and GVHD and the different therapies required, detection of viral load will be effective in patients' survival. 51 recipients of BMT were monitored for 100 days post-BMT during which the samples were collected weekly. The Real-Time PCR was developed for quantitation of CMV viral DNA, using TaqMan tecnnology. For generation of standard curve, UL83 gene from CMV was cloned into a plasmid using a T/A cloning procedure. RQ-PCR assay was preformed in parallel with pp65 antigenemia assay on 415 samples. The results obtained by both techniques were significantly correlated [p < 0.01]. We could detect 13x10[1] -15x10[7] [CMV DNA copies/2x10[5] cells] by RQ-PCR method. About 76% of patients developed at least one episode of CMV reactivation. First positive result of RQ-PCR appeared 13 days earlier than of pp65 antigenemia. After preemptive therapy, 16 days were required to achieve negative result by RQ-PCR. Both assays were highly correlated; however, RQ-PCR was more sensitive than the antigenemia assay. After preemptive therapy, negative results of RQ-PCR were the best indicator to determine the endpoint of treatment and its success

[Multiple carboxylase deficiency]

Multiple carboxylase deficiency is a syndrom in which biotin-dependant carboxylases show diminished activity du to a lack of biotin or autosomal recessively inherited disorders of biotin metabolism. The clinical picture involves the nervous system, skin, respiratory system, digestive system, and immune system. This syndrome is fatal in the absence of prompt diagnosis and treatment with biotin. Authors report the case of 2-month-old child explored for myoclonic seizures that do not respond well to classic anticonvulsivant therapy, hypotonia, skin problems with alopecia, appeared at age of one month. The laboratory examinations showed hyperammonemia and hyperlactacedemia.Multiple carboxylase deficiency was suspected and treatment with biotin [5mg/day] achieved a rapid improvement of the seizures and the skin problems

Evalution of the relationship between vitamin d receptor gene polymorphism [FOK I] and thyroid cancer

Background and purpose: Vitamin D is an antiproliferative agent against cancer cells and regulates cell differentiation. It acts via Vitamin D Receptor [VDR]. The VDR gene contains a Start Colon Polymorphism [SCP] that can be detected with the restriction enzyme Fok I. Previous studies report an association of SCP and some diseases and some suggest that this polymorphism alters VDR function. As no studies so far have reported the association between Fok I genotype in Thyroid cancer, this study is intended to determine the association of Fok I polymorphism of VDR with thyroid cancer risks in Iranian population

Methods and Materials: 58 patients with papillary carcinoma, 13 patients with follicular carcinoma and 82 controls participated in a case-control study. A PCR-RFLP method used to determine VDR gene polymorphism in start codon characterized by the restriction enzyme Fok I. 95% confidence intervals and odds ratio were calculated for testing the relationship between Fok I polymorphism and thyroid cancer

Results: The odds ratio for the Fok I polymorphism on thyroid cancer was 0.39 [95% CI, 0.12-1.27] witch signifies no relationship between this polymorphism and the ris of thyroid cancer

Conclusion: As no relationship was found between this polymorphism and thyroid cancer, other genetic or environmental factors may be considered in thyroid cancer

[Assessment of Wilms tumor gene [WT1] in acute myeloid leukemia patients as a new marker to detect minimal residual disease [MRD]]

WT1 gene encodes a transcription factor that is involved in differentiation and proliferation of hematopoietic precursor cells as well as some other tissues like kidney, ovary, heart, etc. Quantitative assessment of WT1 gene expression is proposed as a useful marker in MRD detection and leukemia management. To assess the relevance of this gene, we analysed peripheral blood mononuclear cells of 62 AML patients [new cases] for the expression level of WT1 mRNA using Real-time quantitative RT-PCR. We followed the analysis up to 3 years, depending on patient availability. This study as a fundamental and applicable one was done cross-sectionnaly. Samples were obtained randomly from the AML patients referred to the BMT center, and selection was based on the diagnostic criteria defined by clinical wards. WT1 expression in MNCs of patients was compared with 24 healthy individuals [K562 cells considered to express WT1 gene equivalent to 10[6]]. Samples for diagnosis showed significantly high levels of WT1 expression [>80%]. After chemotherapy, its expression decreased [diminished about 1-2 log within induction therapy and around 3-4 log after consolidation therapy]. There was a noticeable correlation between the relative expression levels of WT1 and prediction of relapse [lower than gray zone versus higher than]. Patients whose WT1 expression levels remained lower than the gray zone benefit from a better compete remission. On the contrary, 1-6 months prior to overt clinical relapse in 6 patients, their WT1 expression raised to levels upper than gray zone. This study revealed that WT1 is a useful marker for detecting minimal residual disease, assessing chemotherapy effects, and predicting relapse in AML patients

Effects of chimerism on graft versus host disease recurrence and survival after HLA identical marrow transplantation in Iran

The co-existence of recipient's hamatopoietic systems after allogeneic marrow transplantation is called mixed chimerism. Chimerism analysis provides a national method of different conditioning regimens, graft-versus-host disease [GVHD], prophylactic regimens, and cellular therapy to promote engraftment. The association of mixed chimerism with acute graft-versus-host disease [GVHD], disease recurrence, survival, and relapse free survival was investigated in 91 patients 12 and 79 of whom underwent either bone or peripheral blood HLA-identical marrow transplantation respectively. Chimerism was assessed using multiplex amplification of shorty tandem repeats [STR-PCR].cases included thalassemics [19 subjects], AML [29], ALL [20], CMT [18] and others [5].Median age was 21 [age range of 3-50]. There were 38 females [41.8%] and 53 males [58.2%]. Conditioning was busulfan plus cyclophosphamide in 34 patients, busulfan plus fludarabin in 51 patients and busulfan plus fludarabin plus anti-thymocyte globulin in 6 patients. Median of follow up was 13 months. Data was analyzed using SPSS statistical software. On day 30 after transplantation, mixed chimerism [MC] was observed in 15 patients [16.5%], complete donor chimerism [CC] in 72 patients [79%], and no chimerism in 4 patients. The incidence of acute GVHD was significantly lower in mixed chimeras that in complete chimeras [p=0.01] but there was no significant difference in acute GVHD grade [I, II vs. III, IV] between two groups. The incidence of relapse and overall survival were 17.6% and 88.9% respectively showing no significant difference between MC and CC. Relapse free survival was 80.2% and significantly different between two groups. Despite some previous reports, we found no significant difference in survival and relapse rate between MC and CC. Relapse free survival was 80.2% and not significantly different between tw ogroup

Frequency of BCR-ABL fusion transcript in Iranian patients with chronic myeloid leukemia

Reverse transcriptase-polymerase chain reaction [RT-PCR] assay is a useful tool for the detection of fusion transcript resulting from specific chromosomal translocation of the leukemia cells. A specific chromosomal abnormality, the Philadelphia chromosome [Ph], is present in 90% to 95% of CML patients. The aberration results from a reciprocal translocation between chromosome 9 and 22, creating a BCR-ABL fusion gene. There are two major forms of the BCR/ABL fusion gene, involving ABL exon 2, but including different exons of BCR gene. The transcripts b2a2 or b3a2 code for a p210 protein. Another fusion gene leads to the expression of an e1a2 transcript, which codes for a p190 pro-tein. Another, less common fusion genes are b3a3 or b2a3 [p203] and e19a2 [p230]. The incidence of one or other rearrangement in chronic myeloid leukemia [CML] patients varies in different reported series. In general, fusion transcripts are determined individually, a process which is labor intensive in or-der to detect all major fusion transcripts. This study was designed to determine the frequency of different fusion genes in 75 iranian patients with CML. peripheral blood samples were analyzed by multiplex reverse transcriptase poly-merase chain reaction [RT-PCR] from adult patients to detect all types of BCR-ABL transcripts of the t [9:22] and found that all cases were positive for some type of BCR/ABL rearrangement. Most of our patients showed b3a2 fusion gene [62%], while the remaining showed one of the transcripts of b2a2, b3a3, b2a3, e1a2 or coexpression of b3a2 and b2a2. The rate of coexpression of the b3a2 and b2a2 was 5%. In contrast to the other reports, we did not see any coexpression of p210/p190. This may reflect either the sensitivity of the detection techniques used or the possibility of genetic differences be-tween the populations studied. Coexpression may be due to alternative splicing or to phenotypic varia-tion, with clinical course different from classical CML

Quantitative analysis of Epstein - Barr virus DNA load in bone marrow transplant recipients by using real-time PCR

Quantification of Epstein - Barr virus [EBV] in peripheral blood mononuclear cells [PBMNC] of allogenic bone marrow transplant [BMT] recipients is important because EBV-associated posttransplant Lymphoproliferative disease [PTLD] can occur after transplantation due to immunosup-pression therapy. To this end we chose Real-Time PCR using TaqMan probe. For the standard curve, we cloned BALF5 gene of EBV into a plasmid vector. After purification of the EBV-clone and calculation of plasmid copy number, the standard curve was constructed by using serial dilution of the plasmid clone. We were able to detect from 2 to 107 copies per 2x105 PBMNC with wide linear range. The mean EBV DNA copy number was 103.7 copies per 2x105 PBMNC. In this study, No patient of 35 BMT recipients [275 PBMNC samples] developed PTLD during five months follow up post transplant. EBV copy numbers in 22 samples [3 patients] out of 35 BMT recipients were higher than cut off value with symptoms like fever and pulomonary noddes [9%]. The virus load in one patient in the last sample obtained was 72400 copies. We detected low levels of EBV DNA in 20 BMT patients [57/1%]. Real-Time PCR is useful to measure virus load in PBMNC. Detection of EBV in PBMNC samples may be valuable predictive marker to prognosis PTLD. Further studies need to determine ac-curate viral cut off value for treatment patients at risk for PTLD