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Background: Toxoplasma gondii, the obligate intracellular parasite is life threatening in AIDS patients. Diagnosis of toxoplasmosis is based on serological methods especially increasing of IgM and IgG titers, but finding of parasite or its components (antigenemia) may be beneficial method in order to detection of acute toxoplasmosis in immunocompromised patients., Methods: Ninety-four serum samples from HIV positive patients were collected from Sanandaj, Kordistan west of Iran. These patients were lived in Sanandaj of whom 26 were prisoners infected with HIV virus in prison. Toxoplasma gondii antibodies were determined by IgG ELISA. T. gondii antigen was identified by capture-ELISA. PCR was performed on samples with T. gondii antigenemia. CD4+ T cells counts had been determined by flowcytometry and were obtained from records of each patient., Results: Among the examined HIV seropositive individuals, 19.1% (18/94) and 5.3% (5/94) were positive for Toxoplasma-IgG and antigenemia, respectively. Besides, one of the samples was positively detected by PCR method. Mean age of participants was 37.9 ± 9.5 year. Prevalence of IgG antibody and antgenemia was higher in age group of 40-50 years old. The Mean of CD4+ T cells counts of participants (total of HIV+ patients, IgG positive patients and patients with antigenemia) was 699.2 ± 345.2, 655.1 ± 237.9 and 620.2 ± 215.1 respectively., Conclusion: Capture-ELISA and PCR could confirm the T. gondii acute infection in HIV positive patients. For precise diagnosis of acute toxoplasmosis in HIV positive patient, performance of more studies based on more sensitive types of PCR is suggested.

Background: Applying utilization review programs is an appropriate solution to decrease the expenditure, and to increase the efficiency of healthcare systems., Objectives: This paper presents an instrument to measure the level of appropriate admissions and days of stay (DOS) in the pediatric public hospitals of Iran., Materials and Methods: The American version of the Pediatric Appropriateness Evaluation Protocol (PAEP) was modified and adjusted by our group of physicians. They carried out a retrospective study over 100 randomly selected patients. The reliability of the instrument was tested based on the consensus of reviewers using PAEP. In addition, the external validity of the instrument was studied by comparing the evaluations of the reviewers using PAEP and the individual judgments of three clinicians in two public teaching hospitals. Finally, reliability and validity were also calculated by the kappa statistic., Results: With respect to the inter-reliability testing, there was a high level of agreement between reviewers applying the instrument in the admissions criteria and days of stay. Overall agreement was > 77%; also specific inappropriate agreement and specific appropriate agreement were > 61%, and > 72%, respectively. Regarding the validity of the testing, the instrument had a sensitivity of > 0.75, specificity of > 0.67, as well as positive and negative predictive values of > 0.93, and > 0.55, respectively. The kappa statistic for the reviewers (using the instrument for admission and days of stay criteria) were substantial (k = 0.75.5 and 0.71). They were also substantial for clinicians (k = 0.73 and 0.66)., Conclusions: These results showed that the modified PAEP is a reliable and valid instrument to study the appropriateness of admission and days of stay in Iran hospitals. As the developing countries, particularly, Middle East countries have the same status and culture, the result of this study (with minor changes) could be applied in these countries too.

Background: Despite the wide range of investigations on the therapeutic potentials of herbal extracts in cancer cell lines, there is not usually enough data on their probable effects on normal cells. Recently, both anti-tumoral and anti-oxidant properties of Scutellaria litwinowii extracts have been reported on different cancer cell lines. In the present study, the possible protective effects of S. litwinowii root extracts against cytotoxicity and DNA damage induced by hydrogen peroxide (H2O2) on normal cells were investigated using MTT and Comet assays, respectively., Methods: A method of sequential extraction with solvents of different polarities was used to generate methanolic, de-fatted, and dichloromethane fractions. Both MTT and Comet assays were performed here to assess, respectively, changes in cell viability and levels of DNA damage from H2O2. In a pre-treatment regimen, test materials were applied to the cells prior to H2O2 treatment, while in a co-treatment protocol, cells were treated simultaneously with H2O2 and extracts., Results: In the co-treatment studies, only methanolic extract at 1,000 μg/mL (p<0.001) imparted a significant protective effect as shown in the MTT assay. Pre-treatment of cells for 24 h with different concentrations of the test materials did not lead to any significant protection. Results from the Comet assays in the co-treatment studies suggested a significant (p<0.01) protective effect of the test materials against DNA damage caused by H2O2. However, in the pre-treatment studies, only methanolic extract at ≥500 μg/mL showed a protective effect (p<0.01)., Conclusions: Considering the probable high levels of phenolic and flavonoid compounds in the methanolic extract, these compounds may impart the noted protective effects of the S. litwinowii root through the scavenging of free radicals.

Emerging two-dimensional (2D) materials such as transition metal dichalcogenides offer unique and hitherto unavailable opportunities to tailor the mechanical, thermal, electronic, and optical properties of polymer nanocomposites. In this study, we exfoliated bulk molybdenum disulfide (MoS2) into nanoplatelets, which were then dispersed in epoxy polymers at loading fractions of up to 1% by weight. We characterized the tensile and fracture properties of the composite and show that MoS2 nanoplatelets are highly effective at enhancing the mechanical properties of the epoxy at very low nanofiller loading fractions (below 0.2% by weight). Our results show the potential of 2D sheets of transition metal dichalcogenides as reinforcing additives in polymeric composites. Unlike graphene, transition metal dichalcogenides such as MoS2 are high band gap semiconductors and do not impart significant electrical conductivity to the epoxy matrix. For many applications, it is essential to enhance mechanical properties while also maintaining the electrical insulation properties and the high dielectric constant of the polymer material. In such applications, conductive carbon based fillers such as graphene cannot be utilized. This study demonstrates that 2D transition metal dichalcogenide additives offer an elegant solution to such class of problems.

Psyllium has a mucilaginous property that makes it a good candidate to be utilized as an excipient in the preparation of controlled release systems. Various formulations were prepared using theophylline as a model drug and investigated with a view to achieve an ideal slow drug release profile. The addition of hydroxypropyl methylcellulose (HPMC) to psyllium significantly reduced the burst release; however, the percentage of drug release within a 12 h period was too slow and thereby inadequate. This was overcome by the addition of lactose as a hydrophilic filler that enabled a slow release with roughly 80% drug release in 12 h. The inclusion of HPMC within psyllium formulations changed the drug release kinetics from Fickian diffusion to anomalous transport. Granulated formulations demonstrated slower drug release than ungranulated or physical mixture and caused a change in the dissolution kinetics from Fickian diffusion to anomalous transport. Milled granules showed more efficient controlled drug release with no burst release. Milling of the granules also changed the drug release kinetics to anomalous transport. Although psyllium was proved to be a promising polymer to control the drug release, a combination of psyllium-HPMC and formulation processes should be considered in an attempt to achieve a zero-order release.

Paracetamol is a popular over-the-counter analgesic and a challenging model drug due to its poor technological and biopharmaceutical properties such as flowability, compressibility, compactibility and wettability. This work was aimed to alter the crystal habit of paracetamol from elongated to polyhedral-angular via particle engineering whilst maintaining the stable polymorphic form (form I: monoclinic form). The engineered paracetamol crystals obtained in the present investigation showed better technological and biopharmaceutical properties in comparison to the commercial paracetamol. Engineered paracetamol crystals were obtained using antisolvent crystallization technique in the presence of various concentrations (0.1, 0.5 and 1%, w/w) of additives, namely, polyvinyl alcohol (PVA), Avicel PH 102 (microcrystalline cellulose), Brij 58, methylcellulose (MC) and polyethylene glycol having different molecular weights (PEGs 1500, 6000 and 8000). Paracetamols crystallized in the presence of Avicel (or physically mixed with Avicel), Brij 58 and PEG 6000 demonstrated the best compactibility over a range of compaction pressures. Brij-crystallized paracetamol provided the fastest dissolution rate among all the paracetamol batches. Paracetamols crystallized in the presence of PVA or Avicel, or physically mixed with Avicel demonstrated a reduced degree of crystallinity in comparison to the other paracetamols. This study showed that the type, the grade and the concentration of additives could influence the physical stability such as flow, crystallinity and polymorphic transformation of paracetamol, the technological and biopharmaceutical properties of paracetamol. Stable polymorphic form of paracetamol with optimal tableting characteristics can be achieved through particle engineering., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: Toxoplasma gondii is one of the most prevalent parasites of human and warm- blooded animals. Toxoplasmosis is important especially in two groups: pregnant women and immunocompromised patients. If women acquire the primary infection during the pregnancy, it would be life threatening or remains severe disorders for the fetus. This study was performed to evaluate the seroprevalence of T. gondii infection in pregnant women referred to Health Center in Gorgan City, Golestan Province, northern Iran., Methods: Serum samples were collected from pregnant women referred to Health Center in Gorgan City, south eastern Caspian Sea. Anti- Toxoplasma IgG and IgM antibodies were determined by commercially ELISA kits and the relation of infection with socio-demographic and risk factors such as age, education, occupation, cat ownership, soil contact and some other factors was studied., Results: From 555 tested sera of pregnant women referred to Health Center in Gorgan, 39.8% had IgG antibodies against T. gondii and 3.4% were positive for IgM antibodies. A significant correlation was seen between T. gondii infection with age and soil contact., Conclusion: About 60% of pregnant women in Gorgan City are seronegative against T. gondii, so they should considered as at risk persons.

There are limited therapies for severe emphysema. Bronchoscopic treatments of emphysema were introduced to achieve the beneficial physiological changes seen in surgical lung volume reduction; however, at the present time these treatments are mostly aimed at improving quality of life and functional status in patients with emphysema. At this time, none of these minimally invasive approaches have been approved in the United States for treatment of emphysema; however, several novel interventions have demonstrated potential in early-phase clinical trials. We performed a systematic evaluation of the relevant medical literature and present herein an evidence-based review of bronchoscopic treatments for emphysema, with a focus on the current status of this technology in the United States as compared with Europe.

Hydrophilic matrices are a principal technology used for extended release (ER) oral dosage forms and a recent review concluded that their development is currently one of the most important challenges in pharmaceutical research. High molecular weight polyethylene oxides (PEOs) have been proposed as an alternative to hydroxypropylmethylcellulose (HPMC) for the manufacture of controlled release matrix tablets. It is known that PEO's are prone to oxidative degradation which can occur by chain scission and can be catalyzed by metal ions. In this study, we investigated the stability of PEO matrix tablets, of different molecular weight, containing diltiazem hydrochloride, when stored at 40 °C. The results show that there were dramatic increases in the release rate of the diltiazem following storage over only a few weeks, resulting in immediate release profiles after eight weeks, even for the highest molecular weight grade. We employed Gel permeation chromatography (GPC), viscosity and differential scanning calorimetry (DSC) techniques to try and determine the underlying causes of these dramatic shifts in dissolution profiles on storage. The results showed that there were significant decreases in the molecular weight of the PEO's during storage. The second part of the study looked at the addition of three different levels of vitamin E succinate to the tablets. The results clearly demonstrate the ability of the added antioxidant to reverse the significant reductions in molecular weight seen using GPC, viscosity and DSC. Importantly the addition of the antioxidant was able to stabilize the release profile of the diltiazem especially when present at a 1% level. Researchers and those working in pharmaceutical development should be aware of the potential stability risks when making matrix tablets containing PEO's and may wish to consider the addition of an antioxidant to the tablet formulation., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background: Transforming growth factor-β1 (TGF-β1) plays a major role in pleural fibrosis and chemical pleurodesis. Approximately 50% of patients with tunneled pleural catheters (TPCs) for malignant pleural effusions (MPEs) will have autopleurodesis. Pleural TGF-β1 levels have never been measured in the setting of TPC drainage and may predict autopleurodesis., Methods: Pleural fluid was collected prospectively in subjects with MPE and planned TPC insertion. Pleural fluid was collected at time 0 (T0, thoracentecis before TPC placement), time 1 (T1, TPC insertion), and time 2 (T2, 2 wk after TPC insertion). The MPE levels of TGF-β1, vascular endothelial growth factor, and plasminogen activator inhibitor-1 were measured using ELISA. Lactate dehydrogenase levels were also measured at T0. Patients were followed up for autopleurodesis and TPC removal., Results: Nineteen samples were collected from 9 patients with MPE who underwent TPC placement. Three patients died before the study completion. The median level of TGF-β1 of 1243 pg/mL (range, 620 to 4547 pg/mL) at T1 in 6 patients showed a statistically significant rise to 2857 pg/mL (range, 2120 to 7160 pg/mL) after TPC placement (P=0.004, R=0.898). There was no statistically significant increase in TGF-β1 levels from T0 to T1. There was a linear relationship between TGF-β1 and lactate dehydrogenase at T0 (R=0.635). Plasminogen activator inhibitor-1 levels increased from T1 to T2, but was not statistically significant. There was no significant rise in vascular endothelial growth factor from T1 to T2., Conclusions: TPC insertion results in an independent increase in TGF-β1 levels in MPE.

Previous studies in cell lines have suggested a role for melanosomes and related protein trafficking pathways in melanoma drug response. We have investigated the expression of six proteins related to melanosomes and melanogenesis (MITF, GPR143, gp100/PMEL, MLANA, TYRP1, and RAB27A) in pretreatment metastases from melanoma patients (n = 52) with different response to dacarbazine/temozolomide. Microphthalmia-associated transcription factor (MITF) and G-protein coupled receptor 143 (GPR143) showed significantly higher expression in nonresponders compared with responders. The premelanosome protein (gp100/PMEL) has been indicated previously in resistance to cisplatin in melanoma cells, but the expression levels of gp100/PMEL showed no association with response to dacarbazine/temozolomide in our clinical material. We also investigated the effects on chemosensitivity of siRNA inhibition of gp100/PMEL in the MNT-1 melanoma cell line. As expected from the study of the tumor material, no effect was detected with respect to response to temozolomide. However, knockdown of gp100/PMEL sensitized the cells to both paclitaxel and cisplatin. Overall, our results suggest that MITF, and several MITF-regulated factors, are associated with resistance to chemotherapy in melanoma and that different MITF targets can be of importance for different drugs.

Matrices containing PEO fail to provide stable drug release profiles when stored at elevated temperatures for a period of time. The present study aims to stabilize diltiazem HCl release from matrices made from various molecular weights of polyox powders. To this end, various molecular weights of polyox with and without vitamin E (0.25, 0.5 and 1% w/w) were stored at 40°C for 0, 2, 4 and 8 weeks. The aged polyox powders were then mixed with the model drug at a ratio of 1:1 and compressed into tablets. At different time intervals, the aged polyox with vitamin E were taken out of oven and mixed with the drug (1:1 ratio) and compressed into tablets. Dissolution studies showed a significant increase in diltiazem HCl release rate to occur with increased storage time at 40°C ± 1 from tablets made from the aged polyox (no vitamin E). This was as a result of depolymerization of the aged polyox powders as compared to the fresh polyox samples. This was confirmed by differential scanning calorimetry (DSC) which showed a reduction in the melting point of the aged samples. Concentrations of vitamin E as low as 0.25% w/w was able to overcome the quick release of drug from the matrices made from aged polyox powders. DSC traces showed that the melting point of aged polyox samples containing vitamin E remained the same as that of the fresh samples. The presence of vitamin E is essential to stabilize the drug release from polyox matrices containing diltiazem HCl.

Background: Based on recent studies, there are controversial reports on the capacity of tissue cyst forming of Toxoplasma gondii RH strain. In this study, the capacity was evaluated by in vivo and in vitro experiments., Methods: RH strain was subcutaneously inoculated to ten Wistar rats. After one month, their blood, brain, tongue and diaphragm were collected and evaluated by MAT, PCR, pathological and bioassay methods. The parasite was cultivated in the cell monolayer. To change to bradyzoite, the media pH was altered to 6.8. Biological aspect of the bradyzoites was evaluated by incubation in acidic pepsin and it's inoculation in ten BALB/c mice., Results: All rats showed antibodies to Toxoplasma at titers ≥1:320 but no DNA and tissue cyst were detected in the tissues. Following intraperitoneal inoculation of rats' brain homogenate into BALB/c mice, no infection was established in none of the animals. During presence of cell culture, in acid media for a 3-5 days period, cyst-like structures were noticed when they were stained with PAS. The visible bradyzoites in the cysts that were incubated in acid pepsin medium were not able to kill any mice., Conclusion: This study confirmed that Iranian RH strain has lost the potential of tissue cyst forming in rats and bradyzoites cultivated in cell culture lost their resistance to acidic condition, so this strain can be a candidate for future vaccine researches.

Background: Toxoplasma gondii is an important zoonotic pathogen. Vertical transmission of the parasite occurs when females were infected primarily during gestation. This parasite is transmitted to the fetus through the placenta and may cause miscarriage, permanent neurological damage, premature birth and visual impairment. It has been found that mouse is susceptible to Toxoplasma and is particularly an interesting model to the study of congenital infection but whether the entry of T. gondii through vagina route is involved in transmission of the parasite to the placenta and fetus or not., Objectives: The current study aimed to find a route of infection which perhaps carried the parasite under natural conditions in human., Materials and Methods: In the current experimental study, two 6-8 week NMRI female mice were crossed with one male. The pregnant mice were divided into 2 groups: experimental group that was infected by parasite via intra-vaginal (IV) and control group that received the same volume of normal saline via IV. One mouse from each group was killed on the fifth day after infection. The peritoneal fluid, ovary and uterus of mouse samples were taken and divided into two parts. One part used for DNA extraction and the other was kept in formalin and sent for histological study. These steps were repeated seven times and at least 10 mice in each group (case and control) were studied by molecular and histological methods., Results: PCR using DNA extracted from the experimental group showed that the parasite existed in tissues of the uterus and placenta but not in the embryos and peritoneal fluid. PCR using DNA extracted from the control group was negative., Conclusions: Tachyzoite of Toxoplasma and DNA of this parasite were observed in sub mucosa and muscles of the uterus and in the villis of placenta, but not in histological sections of the fetus. Therefore, histological and molecular results were consistent.

Background: To find out different species of helminthes and blood/tissue protozoan parasites of stray dogs and their potential role for transmission of zoonotic species to human in Mashhad, Khorasan Razavi Province, northeast Iran, during 2008-2009., Methods: Totally, 100 stray dogs were selected among Mashhad municipal collection from different sites of the city. Internal organs were examined for any parasites. Helminthes were identified based on morphological characteristics. Smears prepared from peripheral blood as well as liver, spleen and any skin lesion were stained by Giemsa and examined microscopically. Samples obtained from spleen were aseptically cultured in three culture media including NNN, Schneider's Drosophila (HIMEDIA) and RPMI1640 (GIBCO) for isolation of Leishmania spp. The titer of anti-Leishmania and anti-Toxoplasma antibodies were measured by direct agglutination test (DAT) and indirect fluorescent antibody test (IFAT), respectively., Results: 84% of dogs were infected at least with one species of intestinal helminthes. The species of parasites and rate of infection were as follows: Taenia hydatigena (61%), Dipylidium caninum (46%), Mesocestoides lineatus (19%), Echinococcus granulosus (10%), Toxascaris leonina (53%) and Toxocara canis (7%). Anti-Leishmania antibodies were detected by DAT in 8 dogs (8%) at 1:320 titers and higher. Forty seven dogs (47%) showed anti-Toxoplasma titer at 1:10 and 17 (17%) showed titer of ≥1:100. No blood parasites were found in prepared blood smears., Conclusion: The high rate of parasitic infection and presence of zoonotic species especially E. granulosus and T. canis emphasizes the risk of diseases spread in urban areas by stray dogs.

Bone marrow (BM) provides chemoprotection for acute lymphoblastic leukemia (ALL) cells, contributing to lack of efficacy of current therapies. Integrin alpha4 (alpha4) mediates stromal adhesion of normal and malignant B-cell precursors, and according to gene expression analyses from 207 children with minimal residual disease, is highly associated with poorest outcome. We tested whether interference with alpha4-mediated stromal adhesion might be a new ALL treatment. Two models of leukemia were used, one genetic (conditional alpha4 ablation of BCR-ABL1 [p210(+)] leukemia) and one pharmacological (anti-functional alpha4 antibody treatment of primary ALL). Conditional deletion of alpha4 sensitized leukemia cell to nilotinib. Adhesion of primary pre-B ALL cells was alpha4-dependent; alpha4 blockade sensitized primary ALL cells toward chemotherapy. Chemotherapy combined with Natalizumab prolonged survival of NOD/SCID recipients of primary ALL, suggesting adjuvant alpha4 inhibition as a novel strategy for pre-B ALL.

The diagnosis of visceral leishmaniasis (VL) in humans and animal reservoir hosts is difficult, particularly in rural areas where the disease is endemic and laboratory facilities are limited. This study aimed to develop a latex agglutination test (LAT) for the rapid detection of anti-Leishmania antibodies against the A2 antigen derived from the amastigote form as well as those against crude antigens derived from the promastigote form of an Iranian strain of Leishmania (Leishmania) infantum. The A2 antigen (42-100 kDa) was prepared from the amastigote form of L. infantum, purified with electroelution and compared with the crude antigen from the promastigote form of L. infantum. Both antigens showed appropriate intensity reactions, were selected using dot blotting of positive and negative pooled sera and used to sensitize 0.9-μm latex beads. The tests were carried out on sera from 43 symptomatic, human patients with VL confirmed by parasitological examination and direct agglutination test (DAT), 30 healthy controls and 32 patients with other infections but without VL. Canine sera were collected from 63 domestic dogs with VL confirmed using parasitological examinations and DAT and 31 healthy dogs from areas non-endemic for VL. Compared with the controls, human sera from DAT-confirmed patients yielded a sensitivity of 88.4% (95% CI, 82.1-94.5%) and specificity of 93.5% (95% CI, 87.0-99.7%) on A2-LAT (amastigote) when 1:3200 was used as the cut-off titre. A good degree of agreement was found between A2-LAT and DAT (0.914). LAT required 3-5 min to complete, versus the 12-18 h needed for DAT. Compared with the controls, A2-LAT of canine sera from DAT-confirmed cases yielded a sensitivity of 95.2% (95% CI, 95.0-95.4%) and specificity of 100% (95% CI 100%) when 1:320 was used as the cut-off titre. A good degree of agreement was found between A2-LAT and DAT (0.968). Similarly, the sensitivity and specificity of Pro.-LAT (promastigote) was calculated to be 88.4% and 91.9%, respectively for human sera and 96.8% and 90.3%, respectively for canine sera. No statistically significant differences were observed between A2-LAT and Pro.-LAT for the detection of human and canine L. infantum infections. In conclusion, A2-LAT and Pro.-LAT could be used in parallel to screen for L. infantum infections in humans and dogs in areas endemic for VL in Iran., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Past studies have stated that the parasitostatic effect of IFN-γ is most likely due to the starvation of Toxoplasma gondii for tryptophan in the host cell. The aim of this study was to evaluate the direct effect of two new Naphthalene-Sulfonyl-Indole compounds as competitive molecules for tryptophan on viability and infectivity of Toxoplasma tachyzoites. Tachyzoites of RH strain were incubated in various concentrations (25-800 μM) of 1-(naphthalene-2-sulfonyl)-2,3-dihydro-1H-indole and 1-[5-(2,3-dihydro-1H-indole-1-sulfonyl)naphthalene-1-sulfonyl]-2,3-dihydro-1H-indole for 1.5 hours. Then, they were stained by PI and analyzed by FACS. To evaluate the infectivity, 2 × 10(6) tachyzoites exposed to the concentrations mentioned above were intraperitoneally inoculated into five mice from each group. Also, naïve parasites and parasites exposed to DMSO (control) were inoculated in both groups of mice. The LD50 of 1-(naphthalene-2-sulfonyl)-2,3-dihydro-1H-indole was 62 μmol whilst the quantity of 1-[5-(2,3-dihydro-1H-indole-1-sulfonyl)naphthalene-1-sulfonyl]-2,3-dihydro-1H-indole was more than 800 μmol. The infectivity of tachyzoites exposed to both of the compounds preserved and killed mice. No statistical correlation was seen between longevity of mice groups and different doses of the compounds. If we consider a well-organized transporter mechanism for indole compounds in the parasite, thus the designation of an antagonist that has indole groups can assist with the production of new drugs.

To perform IgG avidity immunoblotting assay for detection of acute toxoplasmosis, 100 serum samples were collected from Tehran, Iran. The presence of Toxoplasma-specific IgG and IgM antibodies were checked by commercial Trinity kit. Samples were categorized in acute and chronic phases of Toxoplasma gondii infection according to IgG avidity ELISA. IgG avidity immunoblotting was performed, and antigenic bands with molecular weights of 22, 25, 28, 30, 32, 42, 44, 49, 55, 60, 66, 69, 88, 106, 130 and 157 kDa were recognized as low avidity markers. The most prevalent antigen for low avidity was p22. It is concluded that IgG avidity immunoblotting could distinguish acute and chronic phases of T. gondii infection., (© S. Ali-Heydari et al., published by EDP Sciences, 2013.)

Unlabelled: Genetic lesions such as BCR-ABL1, E2A-PBX1, and MLL rearrangements (MLLr) are associated with unfavorable outcomes in adult B-cell precursor acute lymphoblastic leukemia (B-ALL). Leukemia oncoproteins may directly or indirectly disrupt cytosine methylation patterning to mediate the malignant phenotype. We postulated that DNA methylation signatures in these aggressive B-ALLs would point toward disease mechanisms and useful biomarkers and therapeutic targets. We therefore conducted DNA methylation and gene expression profiling on a cohort of 215 adult patients with B-ALL enrolled in a single phase III clinical trial (ECOG E2993) and normal control B cells. In BCR-ABL1-positive B-ALLs, aberrant cytosine methylation patterning centered around a cytokine network defined by hypomethylation and overexpression of IL2RA(CD25). The E2993 trial clinical data showed that CD25 expression was strongly associated with a poor outcome in patients with ALL regardless of BCR-ABL1 status, suggesting CD25 as a novel prognostic biomarker for risk stratification in B-ALLs. In E2A-PBX1-positive B-ALLs, aberrant DNA methylation patterning was strongly associated with direct fusion protein binding as shown by the E2A-PBX1 chromatin immunoprecipitation (ChIP) sequencing (ChIP-seq), suggesting that E2A-PBX1 fusion protein directly remodels the epigenome to impose an aggressive B-ALL phenotype. MLLr B-ALL featured prominent cytosine hypomethylation, which was linked with MLL fusion protein binding, H3K79 dimethylation, and transcriptional upregulation, affecting a set of known and newly identified MLL fusion direct targets with oncogenic activity such as FLT3 and BCL6. Notably, BCL6 blockade or loss of function suppressed proliferation and survival of MLLr leukemia cells, suggesting BCL6-targeted therapy as a new therapeutic strategy for MLLr B-ALLs., Significance: We conducted the first integrative epigenomic study in adult B-ALLs, as a correlative study to the ECOG E2993 phase III clinical trial. This study links for the first time the direct actions of oncogenic fusion proteins with disruption of epigenetic regulation mediated by cytosine methylation. We identify a novel clinically actionable biomarker in B-ALLs: IL2RA (CD25), which is linked with BCR-ABL1 and an inflammatory signaling network associated with chemotherapy resistance. We show that BCL6 is a novel MLL fusion protein target that is required to maintain the proliferation and survival of primary human adult MLLr cells and provide the basis for a clinical trial with BCL6 inhibitors for patients with MLLr., (©2012 AACR.)

The precise diagnosis of the acute toxoplasmosis in pregnant women and immunocompromsied patients has critical importance. Most of the commercially available assays use the whole Toxoplasma soluble extract as the antigen. However, the assays currently available for the detection of specific anti-Toxoplasma antibodies may vary in their abilities to detect serum immunoglobulins, due to the lack of a purified standardized antigen. The aim of this study was production and evaluation of the usefulness of the recombinant Toxoplasma gondii GRA7 antigen for the serodiagnosis of Toxoplasma gondii IgM and IgG by ELISA. A total of 70 T. gondii IgM positive sera, 74 T. gondii IgG positive sera, and 60 sera from subjects who were not infected with T. gondii were examined. These sera were shown different absorbance values in ELISA test. To control the specificity of the rGRA7 other parasitic diseases, for example, echinococcosis, malaria, leishmaniasis, fascioliasis, and strongyloidiasis were tested of which none showed positive results. Sensitivity and specificity of the generated recombinant IgG ELISA in comparison with commercial ELISA (com ELISA) were 89% and 90%, and the sensitivity and specificity of the generated recombinant IgM ELISA were 96% and 90%, respectively. The results obtained here show that this antigen is useful for diagnostic purposes.

Toxoplasmic encephalitis is caused by reactivation of bradyzoites to rapidly dividing tachyzoites of the apicomplexan parasite Toxoplasma gondii in immunocompromised hosts. Diagnosis of this life-threatening disease is problematic, because it is difficult to discriminate between these 2 stages. Toxoplasma PCR assays using gDNA as a template have been unable to discriminate between an increase or decrease in SAG1 and BAG1 expression between the active tachyzoite stage and the latent bradyzoite stage. In the present study, real-time RT-PCR assay was used to detect the expression of bradyzoite (BAG1)- and tachyzoite-specific genes (SAG1) during bradyzoite/tachyzoite stage conversion in mice infected with T. gondii Tehran strain after dexamethasone sodium phosphate (DXM) administration. The conversion reaction was observed in the lungs and brain tissues of experimental mice, indicated by SAG1 expression at day 6 after DXM administration, and continued until day 14. Bradyzoites were also detected in both organs throughout the study; however, it decreased at day 14 significantly. It is suggested that during the reactivation period, bradyzoites not only escape from the cysts and reinvade neighboring cells as tachyzoites, but also converted to new bradyzoites. In summary, the real-time RT-PCR assay provided a reliable, fast, and quantitative way of detecting T. gondii reactivation in an animal model. Thus, this method may be useful for diagnosing stage conversion in clinical specimens of immunocompromised patients (HIV or transplant patients) for early identification of tachyzoite-bradyzoite stage conversion.

Central airway stenosis is extremely rare in pregnancy and could lead to respiratory and cardiovascular embarrassment, especially at the time of delivery. Initially, patients may not show obvious signs of respiratory difficulty. Early recognition of the disease and anticipatory management of a complicated delivery are very important. We present a pregnant patient with tuberculosis-induced severe tracheobronchial stenosis and discuss the management challenges of her delivery.

Serum samples, 100 in the total number, were collected from different laboratories in Tehran, Iran and tested for anti-Toxoplasma specific IgG and IgM antibodies using indirect immunofluorescent antibody test (IFAT). Using the IgG (chronic) and IgM (acute) positive samples, the IgG avidity test was performed by ELISA in duplicate rows of 96-well microtiter plates. One row was washed with 6 M urea and the other with PBS (pH 7.2), then the avidity index (AI) was calculated. Sixteen out of 18 (88.9%) sera with acute toxoplasmosis showed low avidity levels (AI ≤ 50), and 76 out of 82 (92.7%) sera in chronic phase of infection showed high avidity index (AI>60). Six sera had borderline ranges of AI. The results showed that the IgG avidity test by ELISA could distinguish the acute and chronic stages of toxoplasmosis in humans.

Purpose: Toxoplasma gondii is a widespread protozoan parasite that infects a broad range of warm blooded animals as well as humans. The present study was investigated to evaluate the effects of allium cepa on renal failur in male rats which experimentally infected by Toxoplasma gondii, RH strain., Methods: Wistar male rat (n=40) were allocated into four groups, group one that received tachyzoites of T. gondii (ip) (n=10), group two that received tachyzoites of T. gondii (ip), plus fresh onion juice by gavages method (n=10), group three received just fresh onion juice by gavages method (n=10) and control group (n=10) that received nothing. Animals were kept in standard condition. In 30 day after inducing Toxoplasma infection, 5cc blood was collected for serum protein and TAC levels. Kidney tissues of Rat in whole groups were removed and prepared for apopetosis analysis., Results: Serum protein and kidneys weights were significantly decreased in groups that were infected with T. gondii, in comparison to control and onions groups. Kidneys Apopetosis in toxoplasma group significantly increased in comparison to control group (P<0.05).level of TAC was significantly increased in groups that received onio juice (P<0.05)., Conclusion: This study showed that T. gondii have significantly effect on serum protein and TAC, apopetosis and fresh onion juice returned and treated this harmful effect, so it is suggested that eating of onion is useful in toxoplasma infection.

Tyrosine kinase inhibitors (TKIs) are widely used to treat patients with leukaemia driven by BCR-ABL1 (ref. 1) and other oncogenic tyrosine kinases. Recent efforts have focused on developing more potent TKIs that also inhibit mutant tyrosine kinases. However, even effective TKIs typically fail to eradicate leukaemia-initiating cells (LICs), which often cause recurrence of leukaemia after initially successful treatment. Here we report the discovery of a novel mechanism of drug resistance, which is based on protective feedback signalling of leukaemia cells in response to treatment with TKI. We identify BCL6 as a central component of this drug-resistance pathway and demonstrate that targeted inhibition of BCL6 leads to eradication of drug-resistant and leukaemia-initiating subclones.

Background: Toxoplasma gondii infects a wide range of animals; felines are definitive hosts and other animals including the dogs are intermediate hosts. The aim of this study was to determine the seroprevalence of T. gondii infection in dogs in Tehran, capital of Iran and to investigate possible associated risk factors., Methods: Three hundreds ninety six serum samples were collected during 2007-8 from the dogs. Collected samples were tested using an indirect fluorescent antibody test (IFAT) in dilutions of 1:16 and more. All procedures were carried out in Shahrekord University, Iran. All the data were analyzed using SPSS software, qui square test with confidence interval of 0.95., Results: From evaluated samples, 89 (22.47%) were positive in titers of at least 1:16. further evaluations in other dilutions showed positive results in dilutions of maximum 1:16, 1:32, 1:64, 1:128 and 1:256 in 38, 29, 15, 2 and 5 dogs respectively. Investigation of the role of risk factors showed no sex predisposition while infection rate was significantly higher in dogs older than one year old. Living places were of significant importance; infection rate was significantly higher in stray or guard dogs in compare with household dogs (P<0.05)., Conclusion: Relatively high seroprevalence of T. gondii infection in dogs in Tehran shows high environmental contamination. It is recommended that the dogs with suspected clinical signs be tested for T. gondii infection.

We present results of mutation screening of PRKN gene in 93 Iranian Parkinson's disease (PD) patients with average age at onset (AAO) of 42.2 years. The gene was screened by direct sequencing and by a semi-quantitative PCR protocol for detection of sequence rearrangements. Heterozygous rearrangements were tested by reverse transcription-polymerase chain reaction (RT-PCR). Nine different PRKN mutations were found. One of these, IVS9+1G>A, affects splicing and is novel. Two mutated PRKN alleles were observed in each of 6 patients whose average AAO was 25.7 years. Only 1 patient carried a single mutated allele and his AAO was 41 years. Among patients with AAO of <30 years, 31.3% had two mutated alleles, while only 2.6% with AAO of >30 years carried a PRKN mutation. Analysis of PRKN by RT-PCR led to identification of a novel exon expressed in leukocytes of control and PD individuals. The alternatively spliced transcript if translated would code a protein without a RING Finger 2 domain. Its functional relevance remains to be shown. DJ-I and PINK1 were also screened. Two novel DJ-1 mutations, c.91-2A>G affecting splicing and c.319G>C causing Ala107Pro, were observed among patients with AAO of <31 years, suggesting that PD in a high fraction (>12%) of this group of Iranian patients may be due to mutations in DJ-1. Mutations in PINK1 were not observed. Our results complement previous findings on LRRK2 mutations among Iranian PD patients., (Copyright © 2010 Movement Disorder Society.)

Objective: Oral squamous cell carcinoma (OSCC) is one of the 10 most common malignant tumors and SCC accounts for approximately 94% of all oral malignancies. The risk of malignant transformation in dysplastic lesions is greater than that of normal oral mucosa. The definite roles of mast cells and angiogenesis in OSCC have been under debate. The aim of this study was to compare mast cell count (MCC) and microvessel density (MVD) among normal oral mucosa, oral dysplastic epithelium and low- and high- grade OSCC., Material and Methods: A total of 42 specimens of OSCC (21 high- and 21 low-grade) were collected, along with six normal and 22 dysplastic oral mucosa. The mean MCC and MVD, as well as the correlation between them, were evaluated by immunohistochemical staining., Results: Statistically significant increases in mean MCC and MVD were observed between normal oral mucosa and epithelial dysplasia, normal oral mucosa and OSCC and epithelial dysplasia and OSCC (P < 0.05), but there were no statistically significant differences in MCC and MVD between low- and high-grade OSCC. Also, the Spearman's correlation coefficient showed a significant correlation between MCC and MVD (r = 0.727, P < 0.001)., Conclusions: The significant correlation found between MCC and MVD is in agreement with the idea that mast cells promote tumor progression via upregulation of angiogenesis. MCC and the degree of angiogenesis can potentially be used as indicators of the evolution of SCC from epithelial dysplasia.

BCL6 protects germinal center (GC) B cells against DNA damage-induced apoptosis during somatic hypermutation and class-switch recombination. Although expression of BCL6 was not found in early IL-7-dependent B cell precursors, we report that IL-7Ralpha-Stat5 signaling negatively regulates BCL6. Upon productive VH-DJH gene rearrangement and expression of a mu heavy chain, however, activation of pre-B cell receptor signaling strongly induces BCL6 expression, whereas IL-7Ralpha-Stat5 signaling is attenuated. At the transition from IL-7-dependent to -independent stages of B cell development, BCL6 is activated, reaches expression levels resembling those in GC B cells, and protects pre-B cells from DNA damage-induced apoptosis during immunoglobulin (Ig) light chain gene recombination. In the absence of BCL6, DNA breaks during Ig light chain gene rearrangement lead to excessive up-regulation of Arf and p53. As a consequence, the pool of new bone marrow immature B cells is markedly reduced in size and clonal diversity. We conclude that negative regulation of Arf by BCL6 is required for pre-B cell self-renewal and the formation of a diverse polyclonal B cell repertoire.

The results of mutation screening of 24 exons of LRRK2 in 60 Iranian Parkinson's Disease patients are presented. The Iranian cohort represents a novel population and was notably young (average age at onset of disease: 36.0 years). Fifty sequence variations were found, seventeen of which are novel. Variations considered possibly associated with disease were screened in available family members, 145 additional patients and 220 control individuals. It was surmised that four novel sequence variations (IVS49+178A>G, p.R1725Q, p.Q1823K, and p.D2175H) may be associated with PD status, albeit they may be very rare non-disease associated variations. The four variations were all observed in the heterozygous state in early onset cases. If one or more of the variations do indeed contribute to disease status, their penetrance is expected to be low.

Glaucoma is a heterogeneous group of optic neuropathies that manifests by optic nerve head cupping or degeneration of the optic nerve, resulting in a specific pattern of visual field loss. Glaucoma leads to blindness if left untreated, and is considered the second leading cause of blindness worldwide. The subgroup primary congenital glaucoma (PCG) is characterized by an anatomical defect in the trabecular meshwork, and age at onset in the neonatal or infantile period. It is the most severe form of glaucoma. CYP1B1 was the first gene genetically linked to PCG, and CYP1B1 mutations are the cause of disease in 20-100% of patients in different populations. Here, we report that LTBP2 encoding latent transforming growth factor beta binding protein 2 is a PCG causing gene, confirming results recently reported. A disease-associated locus on chromosome 14 was identified by performing whole genome autozygosity mapping in Iranian PCG families using high density single nucleotide polymorphism chips, and two disease-segregating loss of function mutations in LTBP2, p.Ser472fsX3 and p.Tyr1793fsX55, were observed in two families while sequencing candidate genes in the locus. The p.Tyr1793fsX55 mutation affects an amino acid close to the C-terminal of the encoded protein. Subsequently, LTBP2 expression was shown in human eyes, including the trabecular meshwork and ciliary processes that are thought to be relevant to the etiology of PCG.

We present results of mutation screening of exons 31, 34, 35, 41, and 48 of LRRK2 in 205 Iranian Parkinson's disease patients. Sixteen percent of the cases were familial. Although age was not a factor in patient recruitment, the Iranian cohort was relatively young (average age at onset of disease: 48.9 years). A notably high male to female ratio (2.96:1) and earlier age at onset (by 2.9 years) in men were observed. A known disease-associated variation, c.C4321T causing R1441C, and IVS31 + 3A > G, a variation that may be associated, were observed. Therefore, disregarding IVS31 + 3A > G, disease status in at least 0.5% of our young cohort and in 3.5% of the familial cases was associated with a mutation in the five exons of LRRK2 screened. Interestingly, the variation causing p.G2019S was not observed., ((c) 2009 Movement Disorder Society.)

Background: PLA2G6 mutations are known to be responsible for infantile neuroaxonal dystrophy (INAD) and neurodegeneration with brain iron accumulation (NBIA). In addition, novel mutations in PLA2G6 have recently been associated with dystonia-parkinsonism in two unrelated consanguineous families., Methods: Direct sequencing analysis of the PLA2G6 gene., Results: Here, we report the segregation of R632W with disease in an Iranian consanguineous dystonia-parkinsonism pedigree. The identical mutation was previously observed in a patient affected with NBIA., Conclusion: We conclude that different and even identical PLA2G6 mutations may cause neurodegenerative diseases with heterogeneous clinical manifestations, including INAD, NBIA and dystonia-parkinsonism.

Robust SNP genotyping technologies and data analysis programs have encouraged researchers in recent years to use SNPs for linkage studies. Platforms used to date have been 10 K chip arrays, but the possible value of interrogating SNPs at higher densities has been considered. Here, we present a genome-wide linkage analysis by means of a 500 K SNP platform. The analysis was done on a large pedigree affected with Parkinsonian-pyramidal syndrome (PPS), and the results showed linkage to chromosome 22. Sequencing of candidate genes revealed a disease-associated homozygous variation (R378G) in FBXO7. FBXO7 codes for a member of the F-box family of proteins, all of which may have a role in the ubiquitin-proteosome protein-degradation pathway. This pathway has been implicated in various neurodegenerative diseases, and identification of FBXO7 as the causative gene of PPS is expected to shed new light on its role. The performance of the array was assessed and systematic analysis of effects of SNP density reduction was performed with the real experimental data. Our results suggest that linkage in our pedigree may have been missed had we used chips containing less than 100,000 SNPs across the genome.

Toxoplasms gondii was detected in sera and urine of acutely infected mice. Animals were inoculated intraperitoneally with tachyzoites of T. gondii, RH strain. Ten animals were killed every day from day 1 up to day 7 post infection. Urine and sera of animals were collected and stored at -20 degree C until use. PCR performed by B1 gene amplification. Toxoplasma was detected in sera from 3 days post infection and in urine from 5 days post infection. No parasite was detected in control group.

Background: Human exposure to dioxins and furans through diet and other sources has been of concern for many years. One specific concern, related to exposure in women's health, is the possible link to endometriosis. Although there are differences in opinion about this link, the concern from the public is real. Congressional interest has prompted investigations to determine the amounts of dioxins and furans present in feminine hygiene products available within the United States., Methods: Tampon samples were analyzed via Gas Chromatography/High Resolution Mass Spectrometry (GC/HRMS) using a Micromass AutoSpec Ultima high resolution mass spectrometer at 10,000 mass resolution. As data were confirmed and quantified using direct isotope dilution, only the 17 2,3,7,8-chlorine-containing dioxin and furan concentrations were calculated from these analyses., Results: A total toxic equivalence (TEQ), using the World Health Organization's toxic equivalency factor (TEF) values, was calculated for each sample. The calculated TEQs for samples were not statistically different from those of the calculated TEQs using the average limit of detection (LOD) values., Conclusions: Data show results similar to those reported by DeVito and Schecter (Environ Health Perspect 2002;110:23) in that most of the dioxins and furans were below the detection limit or estimated detection limits (EDLs).

Three patients are described in whom purulent pericarditis arose as a complication of hydatid disease. In one patient a myocardial cyst ruptured into the pericardial cavity and subsequently became superinfected, probably from a coexisting bronchopneumonia. This patient also had widespread intravascular dissemination of germinal membrane. Two patients had echinococcal cysts of the liver, which became infected following surgery. This led to formation of a fistula between the hepatic hydatid and the pericardium, resulting in purulent pericarditis. The cases illustrate the protean nature of echinococcosis, especially the cardiac form, and emphasize the difficulty of recognizing purulent pericarditis as a complication of other conditions.