Cloning and expression of cyclophilin from Platanus orientalis pollens in Escherichia coli

Background:

Allergy is a clinical disorder affecting the human population with wide geographical distribution. Platanus orientalis (P. orientalis) trees are planted in many countries and their pollen causes allergic reactions.Cyclophilin has recently been identified as one of the most important allergens of P. orientalis pollen. We aimed to clone and purify this allergen in Escherichia coli for further studies and therapeutic and diagnostic purposes for allergy to P. orientalis.

Methods:

RNA was extracted from P. orientalis. A full-length fragment encoding cyclophilin was prepared by polymerase chain reaction amplification of the first-strand cDNA synthesized from P. orientalis RNA. The cDNA was inserted into the pET32b (+) vector, and the construct transformed into E. coli Top10 and BL21 cells. The expressed protein was purified by the CuSO4 method.

Results:

The cDNA for the cyclophilin of P. orientalis pollen was cloned, and a specific reactivity of recombinant cyclophin was confirmed by immunoblotting using sera from patients allergic to P. orientalis pollen.

Conclusion:

The recombinant cyclophilin has a potential for immunologic assays for evaluation of allergy to P. orientalis pollen.Key Words: Allergy; Recombinant allergen; Cyclophilin, Escherichia coli, Platanus orientalis, Pollen, Cloning     

Comparison of Carnoy's solution and 96% ethyl alcohol fixation in bloody Pap smears

OBJECTIVE: To compare 2 methods of fixation in bloody Pap smears with Carnoy's solution and 96% ethyl alcohol. STUDY DESIGN: After observation of contact bleeding, 2 samples were prepared from cervical cells with conventional Pap smear. One sample was fixed in 96% ethyl alcohol and another sample was fixed in Carnoy's solution. RESULTS: Of 450 slides, 410 were selected for study. In study of cell adequacy, diagnosis of squamous cells and glandular cells was better in Carnoy's-fixed slides. Blood contamination of slides was reduced in Carnoy's-fixed slides (13.85% vs. 49.51%), and clearance of slides was increased in Carnoy's-fixed slides. Diagnosis of inflammatory cells and pathogenic microorganisms in was increased in Carnoy's-fixed slides, but no difference was seen in diagnosis of epithelial cell and glandular cell abnormalities. CONCLUSION: Carnoy's solution can be used as an effective fixative in bloody smears in conventional Pap tests.     

Saccharomyces cervisiae as an Efficient Carrier for Delivery of Bioactives: a Review

Food Biophysics - Recently, there has been growing interest in usingnatural vesicles for encapsulation of variousfood-grade ingredients. The structure ofyeast cells, along with their presence in...     

Variation in calcification rate of Acropora downingi relative to seasonal changes in environmental conditions in the northeastern Persian Gulf

There is a strong interest in understanding how coral calcification varies with changing environmental conditions, especially given the projected changes in temperature and aragonite saturation due to climate change. This study explores in situ variation in calcification rates of Acropora downingi in the northeastern Persian Gulf relative to seasonal changes in temperature, irradiance and aragonite saturation state (Ω _arag). Calcification rates of A. downingi were highest in the spring and lowest in the winter, and intra-annual variation in calcification rate was significantly related to temperature (r ² = 0.30) and irradiance (r ² = 0.36), but not Ω _arag (r ² = 0.02). Seasonal differences in temperature are obviously confounded by differences in other environmental conditions and vice versa. Therefore, we used published relationships from experimental studies to establish which environmental parameter(s) (temperature, irradiance, and/or Ω _arag) placed greatest constraints on calcification rate (relative to the maximum spring rate) in each season. Variation in calcification rates was largely attributable to seasonal changes in irradiance and temperature (possibly ~57.4 and 39.7% respectively). Therefore, we predict that ocean warming may lead to increased rates of calcification during winter, but decelerate calcification during spring, fall and especially summer, resulting in net deceleration of calcification for A. downingi in the Persian Gulf.

     

Synthesis,cytotoxicity assessment,and interaction and docking of novel palladium(II) complexes of imidazole derivatives with human serum albumin

Imidazole analogs are the agents that attract both bioinorganic chemist and drug designer. Numerous methods have been proposed for synthesis of imidazole derivatives. In this study, a series of heterocyclic system with p-conjugated system such as 2-aryl-imidazo[4,5-f][1,10]phenanthroline analogs were synthesized. Then, three new palladium(II) complexes containing 2-(Furan-2-yl)-1H-Imidazo[4,5-f][1,10]Phenanthroline (FIP) and 2-(thiophen-2-yl)-1H-imidazo[4,5-f][1,10]phenanthroline (TIP) ligands were synthesized. The structures of the compounds, [Pd(Phen)(TIP)](NO₃)₂, [Pd(Phen)(FIP)](NO₃)₂, and [Pd(FIP)₂]Cl were determined by spectroscopic methods and elemental analysis. Biological activity of the complexes synthesized was assessed against chronic myelogenous leukemia cell line, K562. Also, the interactions of human serum albumin with complexes were investigated using isothermal titration in the Tris buffer, pH 7.4. According to the results obtained, it was found that there is a set of six binding sites for these complexes on HSA with positive cooperativity in the binding process. Docking technique was also applied to confirm the experimental results. The results showed that smaller complexes have higher interaction affinity.     

Ant colony based constrained workflow scheduling for heterogeneous computing systems

The problem of constrained workflow scheduling on heterogeneous computing systems has been of major interest in the recent years. The user requirements are described by defining constraints on the workflow makespan and/or its execution cost. The uncertainty in the activity execution path and the dynamicity in the resource workload may cause some run-time changes of the makespan or cost. To prohibit run-time constraint violation, the system needs robust schedules. In this paper, probability of violation (POV) of constraints is proposed as a criterion for the schedule robustness. An ant colony system is then used to minimize an aggregation of violation of constraints and the POV. Simulation results on real world workflows show the effectiveness of the proposed method in finding feasible schedules. The results also indicate that the proposed method decreases the POV, as well as the expected penalty at run-time.     

A Novel Mitochondrial Heteroplasmic C13806A Point Mutation Associated with Iranian Friedreich’s Ataxia

Friedreich’s ataxia (FRDA) is an autosomal recessive neurodegenerative disorder caused by decreased expression of the protein Frataxin. Frataxin deficiency leads to excessive free radical production and dysfunction of chain complexes. Mitochondrial DNA (mtDNA) could be considered a candidate modifier factor for FRDA disease, since mitochondrial oxidative stress is thought to be involved in the pathogenesis of this disease. It prompted us to focus on the mtDNA and monitor the nucleotide changes of genome which are probably the cause of respiratory chain defects and reduced ATP generation. We searched about 46% of the entire mitochondrial genome by temporal temperature gradient gel electrophoresis (TTGE) and DNA fragments showing abnormal banding patterns were sequenced for the identification of exact mutations. In 18 patients, for the first time, we detected 26 mtDNA mutations; of which 5 (19.2%) was novel and 21 (80.8%) have been reported in other diseases. Heteroplasmic C13806A polymorphisms were associated with Iranian FRDA patients (55.5%). Our results showed that NADH dehydrogenase (ND) genes mutations in FRDA samples were higher than normal controls (P < 0.001) and we found statistically significant inverse correlation (r = −0.8) between number of mutation in ND genes and age of onset in FRDA patients. It is possible that mutations in ND genes could constitute a predisposing factor which in combination with environmental risk factors affects age of onset and disease progression.     

PCR-based detection and eradication of mycoplasmal infections from various mammalian cell lines: a local experience

A total of 200 cell lines including different human, monkey, mice, hamster and rat cell types were examined for mycoplasma infection status. PCR assay using generic-specific universal primers showed that 40 (20%) of the cell lines are contaminated with mycoplasma. Employment of species-specific primers within these infected cell lines revealed infection with M. hyorhinis (42.5%), M. fermentas (37.5%), M. arginini (37.5%), M. orale (12.5%) and A. laidlawii (7.5%). A number of the cultures were coinfected with 2 or 3 different species. Contaminated samples were treated with BM-Cyclin, Ciprofloxacin and mycoplasma removal agent (MRA). Mycoplasma eradication was subsequently checked by PCR following 2 weeks continuous culture of treated cells in antibiotic free culture medium. Mycoplasmal infections were eradicated in 100, 70 and 42% of infected cell lines when the samples were treated with BM-Cyclin, MRA and Ciprofloxacin, respectively. However, 12% (BM-Cyclin), 62.5% (MRA) and 82.5% (Ciprofloxacin) of mycoplasma regrowth was observed 4 months after the treatment. Notably, the risk of spontaneous culture death was 17.5, 12.5 and 0% for BM-Cyclin, MRA and Ciprofloxacin, respectively.