[Effect of spirituality therapy on the resilience of women with breast cancer in Tehran, Iran]

Background and Objective: Spirituality refers to considering the cultural-religious beliefs of the people in the therapeutic process and taking into account the transcendental dimension of the clients who lead them to the transcendental source. This study was conducted to determine the effect of spirituality therapy on the resilience of women with breast cancer in Tehran, Iran

Methods: This quasi-experimental study was performed on 30 women with breast cancer referred to the oncology and chemotherapy clinic in Tehran, Iran during 2016. Subjects were selected by available sampling method and then non-randomly divided into two groups of 15 intervention and control groups. The intervention group was subjected to psychological intervention during 11 sessions of 60 minutes and the control group did not receive intervention. Patients completed the Conner and Davidson resiliency questionnaire [2003] before and after the end of the treatment period

Results: The mean and standard deviation of the resiliency score of the intervention and control groups in the beginning of the study were 3.64+/-0.22 and 3.77+/-1.13, respectively. This rate was 4.30+/-0.41 and 3.68+/-0.1 in patients in intervention and control groups, respectively [P<0.05]

Conclusion: Spiritual therapy intervention increased the resiliency of women with breast cancer

Quality control of widely used therapeutic recombinant proteins by a novel real-time PCR approach

Background: Existence of bacterial host-cell DNA contamination in biopharmaceutical products is a potential risk factor for patients receiving these drugs. Hence, the quantity of contamination must be controlled under the regulatory standards. Although different methods such as hybridization assays have been employed to determine DNA impurities, these methods are labor intensive and rather expensive. In this study, a rapid real-time PCR test was served as a method of choice to quantify the E. coli host- cell DNA contamination in widely used recombinant streptokinase [rSK], and alpha interferon [IFN-alpha] preparations

Methods: A specific primer pair was designed to amplify a sequence inside the E. coli 16S rRNA gene. Serial dilu ons of DNA extracted fromE. coli host cells, along with DNA extracted from Active Pharmaceutical Ingredients of rSK, and IFN-alpha samples were subjected to an optimized real-time PCR assay based on SYBR Green chemistry

Results: The test enabled us to detect a small quantity of genomic DNA contamination as low as 0.0002 pg in recombinant protein-based drugs. For the first time, this study showed that DNA contamination in rSK and IFN-alpha preparation manufactured in Pasteur Institute of Iran is much lower than the safety limit suggested by the US FDA

Conclusion: Real-time PCR is a reliable test for rapid detection of host-cell DNA contamination, which is a major impurity of therapeutic recombinant proteins to keep manufacturers' minds on refining drugs, and provides consumers with safer biopharmaceuticals

[Detection of HCMV infection in glioma brain tumors]

Human cytomegalovirus [HCMV] is a beta-herpesvirus that causes persistent infection in humans, as well as severe disease in fetuses and immunocompromised individuals. Although HCMV is not currently causally implicated in human cancer, emerging evidence suggests that HCMV infection may be specifically associated with malignancies such as gliomas. Gliomas are one of the most common brain tumors that affect humans. It is classified into four grades. In this study, we have developed and used a real-time PCR method for the detection and diagnosis of HCMV infection in glioma brain tumor samples. Paraffin-embedded tumor samples were chosen from patients who referred to Imam Khomeini Hospital Neurosurgery Ward. DNA was extracted from paraffinembedded tissues by a DNA extraction kit. After designing specific primers for the HCMV US28 region, a real-time PCR method was developed for detection of HCMV US28. The results of qualitative real-time PCR on 4/18 patients [22.2%] were positive. Two patients with positive HCMV results died. This is the first study that has monitored HCMV genes in samples from glioma patients in Iran. Considering the results of this study and controversies associated with other studies, a more comprehensive study using this and other diagnostic methods is suggested.

[Transfection of a hepatoma cell line by cloned HBV DNA and evaluation of transcription and expression of viral-specific markers and interferon stimulated genes]

Despite availability of an effective vaccine against hepatitis B virus [HBV], the global prevalence of this virus infection has not diminished significantly. Contrary to numerous other human viruses, HBV does not have the ability to propagate in cell culture. However, infectious virus has been produced by transfection of human hepatoma cells with plasmids that contain full length HBV genome. Generation and optimization of appropriate cell culture systems can help us in demonstrating the quality of genome replication by PCR as well as expression of surface antigen secretion. Interferon stimulating genes [ISGs] are usually produced in response to interferon and can be determined as a measure of response to IFN-therapy. Therefore, in pharmacological studies, in addition to assessing the effects of a medicine on viral determinants of replication, its' effects on stimulation of various ISGs, as indicators of innate immune responses, can be achieved. In this study, we transfected the Huh-7 hepatoma cell line with pCH-9/3091. HBsAg production and viral mRNA transcription were subsequently evaluated. In this system, by using ISGs-specific primers, the ISG mRNAs recognition method was optimized and utilized. Huh-7 cells supported HBV replication. The peak HBsAg secretion was observed at 72 h post-transfection. By using designed primers for the S and pg/pC regions, transcription and genome replication of the virus was shown. RT-PCR results for ISG production by transfected cells showed no role for HBV in enhancement of ISGs levels in Huh-7 cells. The results indicated that this system can be used for functional studies of HBV-specific genes as well as assessment of the effects of new drugs or new vaccines. In addition, it may be used to study the mechanisms of drug resistance that have resulted in difficulties in response to HBV antivirals, including IFN-alpha

Rapid low-cost detection of hepatitis C virus RNA in HCV-infected patients by real-time RT-PCR using SYBR green I

We intend to design and validate a low-cost assay for the detection of hepatitis C virus [HCV] RNA using rapid-cycle RT-PCR. The procedure is performed in a closed system with little risk of contamination allowing PCR and product identification to be performed within one or two hours. A SYBR Green-based real-time RT-PCR for rapid detection of HCV. Amplicon synthesis was monitored continuously by SYBR Green I, which binds to double stranded DMA during PCR. The PCR products were identified by melting curve analysis. Standard sera with known concentrations of HCV RNA and 150 clinical samples were used to validate our assay. The minimum detection level of our assay was less than 50 ID/mL. The results on 100 plasma samples were comparable with commercial assays. This method is useful for rapid qualitative detection of HCV infection and particularly suitable for routine diagnostic applications

inverse relation between CagA + strains of helicobacter pylori infection and risk of erosive GERD

The aim of this study is investigating the association of Helicobacter pylori H. pylori infection and its cytotoxic-associated gene A cagA strain with reflux esophagitis. In a case-control setting May 2005-2006, patients with reflux esophagitis case group were compared with age and gender matched people suffering from symptoms of gastroesophageal reflux disease with normal upper gastrointestinal endoscopic findings control group in Imam Khomeini Hospital, Tabriz, Iran. The rates of H. pylori and its cagA positive infections were separately compared between the 2 groups and the subgroups with different severity of reflux esophagitis. Ninety-two and 93 patients were enrolled in the case and the control groups. The rate of H. pylori infection was insignificantly lower in the case group 81.5% versus 87.10%, p=0.29, odd ratio 0.654, 95% Confidence interval [CI] 0.293 to 1.495. The CagA positive infections were found significantly more frequent in the control group 59.1% versus 40.2%, p=0.01, odd ratio 0.465, 95% CI 0.258 to 0.836. There was no significant difference between the severity subgroups of the disease for H. pylori p=0.30 or cagA positive infection rates p=0.40. The CagA positive strains might have a protective effect against reflux esophagitis

use of a sensitive RT-Nested PCR method for detection of Hepatitis C virus

Hepatitis C virus is the major cause of viral hepatitis and its diagnosis in suspected specimens is of great importance. The risk of transfusion- transmitted virus infection is primarily the result of failure in serological screening tests to detect recently infected donors in the pre-seroconversion window period of infection. Therefore, sensitive and accurate diagnosis of HCV prior to antibody production to reduce window period is necessary. In the present study, a sensitive and specific RT-Nested PCR method for detection of a conserved HCV 5 UTR sequence was developed. Two pairs of primers for amplification of the target sequence in two rounds of PCR were selected. The developed RT- Nested PCR assay was performed on HCV-antibody confirmed positive samples as well as negative controls and standard samples. In order to compare the results, One Step RT-PCR kit was used in this study. 25 HCV-positive plasma samples whose positivity were confirmed by ELISA and Western Blot tests, also as well as 10 fold dilutions of a high viral load plasma sample obtained from a HCV-positive patient as standard samples and 25 negative control plasmas from healthy blood donors were collected and tested by this assay. In all of positive samples a 175bp band was observed on agarose gel electrophoresis, but no band could be detected in negative control plasma. Results from developed RT-PCR assay and One Step RT-PCR kit showed a good correlation. According to the results of this study, the developed RT-Nested PCR assay has a good sensitivity and specificity for diagnosis of HCV infection. It has the advantage of viral genome detection prior to seroconversion and can be used to detect HCV infection during window period

Monitoring pyrethroid insecticide resistance in major malaria vector anopheles culicifacies: comparison of molecular tools and conventional susceptibility test

Anopheles culicifacies is a main malaria vector in southeastern part of Iran, bordering Afghanistan and Pakistan. So far, resistance to DDT, dieldrin, malathion and partial tolerance to pyrethroids has been reported in An. stephensi, but nothing confirmed on resistance status of An. culicifacies in Iran. In current study, along with WHO routine susceptibility test with DDT [4%], dieldrin [0.4%], malathion [5%], permethrin [0.25%], lambadacyhalothrin [0.1%], and deltamethrin 0.025, we cloned and sequenced segment VI of domain II [SII6] in voltage-gated sodium channel [vgsc] gene of An. Culicifacies specimens collected in Sistan and Baluchistan province [Iran]. A 221-bp amplified fragment showed 91% and 93% similarity with exon I and exon II of An. gambiae. The size of intron II in An. culicifacies is 62 bp, while in An. gambiae is 57 bp. The major difference within An. culicifacies specimens and also with An. gambiae is in position 29 of exon I, which led to substitution of Leu to His amino acid. This data will act as first report on partial sequence of vgsc gene and its polymorphism in An. culicifacies. A Leu to His amino acid substitution detected upstream the formerly known knockdown resistance [kdr] mutation site could be an indication for other possible mutations related to insecticide resistance. However, the result of WHO susceptibility test carried out in Baluchistan of Iran revealed a level of tolerance to DDT and dieldrin, but almost complete susceptibility to pyrethroids in An. culicifacies. We postulate that the molecular diagnostic tool developed for detection and identification of kdr-related mutations in An. culicifacies, could be useful in monitoring insecticide resistance in Iran and neighbouring countries such as Pakistan and Afghanistan. A phylogenetic tree also constructed based on the sequence of exon I and II, which readily separated An. culicifacies populations from An. stephensi, An. fluviatilis and An. gambiae

Detection and genotyping of hepatitis D virus from HBsAg positive patients in Iran using RT-PCR

Hepatitis Delta virus [HDV] is a degenerate RNA virus or virusoid and a satellite of Hepatitis B virus [HBV]. Three distinct genotypes are described for HDV; genotype I is distributed worldwide but other genotypes appear to be more restricted geographically. In the present study, an RT-nested PCR method was set up to detect delta infection from serum samples. Moreover, the target amplified sequences corresponding to the Hepatitis delta antigen [HDAg] C-termini were used for genotyping. The results showed that 63.6% [23 of 36] of [HDAb] positive serum samples [as determined by ELISA] were also positive for HDV-RNA. Sequencing and phylogenic analysis of three Iranian HDV isolates revealed the most homology [93%] with an Italian isolate indicating a close relationship and probably a common origin for these isolates

Development of a sensitive quantitative competitive PCR assay for detection of human cytomegalovirus DNA

Accurate and rapid diagnosis of human cytomegalovirus [HCMV] disease in immunocompromised patients has remained as a challenge. Quantitative competitive PCR [QC-PCR] methods for detection of HCMV in these individuals have improved the positive and negative predictive values of PCR for diagnosis of HCMV disease. In this study we used QC-PCR assay, using a co-amplified DNA standard, to quantitate the HCMV glycoprotein B [gB] gene in different samples. A DNA internal standard [IS] was designed by replacing HCMV primer binding site at 5' ends of primers that amplifies a 156-bp fragment of lambda genome, and a 200 bp amplicon was produced. Two DNA fragments of 257 bp wild type and 200-bp [IS] were co-amplified with the same oligonucleotide primer sets, analyzed by gel electrophoresis and used for construction of a standard curve. From this, the copy number of the gB gene present in different samples could be determined. Co-amplification with 1,000 copies of IS, allowed quantitation of 10-100,000 of HCMV DNA in a single PCR. This rapid assay avoids using radioactive components and other less efficient quantitative systems. It has the potential for early identification of patients at high risk of development of HCMV disease, and is useful for therapeutic monitoring