The immunotoxin activity of exotoxin A is sensitive to domain modifications.

Immunotoxins are a class of recombinant proteins which consist of an antibody and a part of a bacterial or herbal toxin. Immunotoxins containing Pseudomonas aeruginosa exotoxin A (PEA) have been found to be very applicable in clinical trials. Many obstacles such as solubility and absorbency reduce their usability in solid tumors. The current study aims to overcome the mentioned barriers by addition and removal of functional and non-functional domains with a structural approach. In the experimental section, we took advantage of molecular dynamics simulations to predict the functionality of candidate immunotoxins which target human HER2 receptors and confirmed our findings with in vitro experiments. We found out when no changes were made to domain II of PEA, addition of solubilizing domains to immunotoxins would not reduce their targeting and anti-tumor activity, while increasing the yield of expression and stability. On the other side, when we replaced domain II with eleven amino acids of furin cleavage site (FCS), the activity of the immunotoxin was mainly affected by the FCS neighboring domains and linkers. A combination of seven beneficial point mutations in domain III was also assessed and reconfirmed that the toxicity of the immunotoxin would be reduced dramatically. The obtained results indicate that the addition or removal of domains cannot depict the activity of immunotoxins and the matter should be assessed structurally in advance.

Russian olive (Elaeagnus angustifolia) as a herbal healer.

Introduction:Elaeagnus spp. is one in the family of riparian trees growing near the rivers or water corridors. In this family, Elaeagnus angustifolia (Russian olive) is famous because of its medical applications. Methods: A comprehensive review was performed to extract the related data from published literature. Results: Traditionally, it has been used as an analgesic, antipyretic and diuretic herbal medicine. A large number of compounds have been derived from Russian olive and made this plant a source of flavonoids, alkaloids, minerals and vitamins. Although the purpose of most studies is to use this plant for preparation of herbal medicines and as an ingredient for drug formulation, there is no available drug dosage form commercially. Conclusion: This review aimed to provide the most important documentary information on the active components of Elaeagnus spp. and their relation to the pharmacological properties and compare them with reported medicinal effects.

Two novel mutations in CYP11B1 and modeling the consequent alterations of the translated protein in classic congenital adrenal hyperplasia patients.

Mutations in the 11ß-hydroxylase (CYP11B1) gene are the second leading cause of congenital adrenal hyperplasia (CAH), an autosomal recessive disorder characterized by adrenal insufficiency, virilization of female external genitalia, and hypertension with or without hypokalemic alkalosis. Molecular analysis of CYP11B1 gene in CAH patients with 11ß-hydroxylase deficiency was performed in this study. Cycle sequencing of 9 exons in CYP11B1 was performed in 5 unrelated families with 11ß-hydroxylase deficient children. Three-dimensional models for the normal and mutant proteins and their affinity to their known substrates were examined. Analysis of the CYP11B1 gene revealed two novel mutations, a small insertion in exon 7 (InsAG393) and a small deletion in exon 2 (DelG766), and three previously known missense mutations (T318M, Q356X, and R427H). According to docking results, the affinity of the protein to its substrates is highly reduced by these novel mutations. DelG766 has more negative impact on the protein in comparison to InsAG393. The novel mutations, InsAG393 and DelG766, change the folding of the protein and disrupt the enzyme's active site as it was measured in the protein modeling and substrate binding analysis. Molecular modeling and sequence conservation were predictive of clinical severity of the disease and correlated with the clinical diagnosis of the patients.

Artificial Neural Networks Analysis Used to Evaluate the Molecular Interactions between Selected Drugs and Human Cyclooxygenase2 Receptor.

OBJECTIVE(S): A fast and reliable evaluation of the binding energy from a single conformation of a molecular complex is an important practical task. Artificial neural networks (ANNs) are strong tools for predicting nonlinear functions which are used in this paper to predict binding energy. We proposed a structure that obtains binding energy using physicochemical molecular descriptions of the selected drugs. MATERIAL AND METHODS: The set of 33 drugs with their binding energy to cyclooxygenase enzyme (COX2) in hand, from different structure groups, were considered. 27 physicochemical property descriptors were calculated by standard molecular modeling. Binding energy was calculated for each compound through docking and also ANN. A multi-layer perceptron neural network was used. RESULTS: The proposed ANN model based on selected molecular descriptors showed a high degree of correlation between binding energy observed and calculated. The final model possessed a 27-4-1 architecture and correlation coefficients for learning, validating and testing sets equaled 0.973, 0.956 and 0.950, respectively. CONCLUSION: RESULTS show that docking results and ANN data have a high correlation. It was shown that ANN is a strong tool for prediction of the binding energy and thus inhibition constants for different drugs in very short periods of time.

Use of spectroscopic, zeta potential and molecular dynamic techniques to study the interaction between human holo-transferrin and two antagonist drugs: comparison of binary and ternary systems.

For the first time, the binding of ropinirole hydrochloride (ROP) and aspirin (ASA) to human holo-transferrin (hTf) has been investigated by spectroscopic approaches (fluorescence quenching, synchronous fluorescence, time-resolved fluorescence, three-dimensional fluorescence, UV-vis absorption, circular dichroism, resonance light scattering), as well as zeta potential and molecular modeling techniques, under simulated physiological conditions. Fluorescence analysis was used to estimate the effect of the ROP and ASA drugs on the fluorescence of hTf as well as to define the binding and quenching properties of binary and ternary complexes. The synchronized fluorescence and three-dimensional fluorescence spectra demonstrated some micro-environmental and conformational changes around the Trp and Tyr residues with a faint red shift. Thermodynamic analysis displayed the van der Waals forces and hydrogen bonds interactions are the major acting forces in stabilizing the complexes. Steady-state and time-resolved fluorescence data revealed that the fluorescence quenching of complexes are static mechanism. The effect of the drugs aggregating on the hTf resulted in an enhancement of the resonance light scattering (RLS) intensity. The average binding distance between were computed according to the forster non-radiation energy transfer theory. The circular dichroism (CD) spectral examinations indicated that the binding of the drugs induced a conformational change of hTf. Measurements of the zeta potential indicated that the combination of electrostatic and hydrophobic interactions between ROP, ASA and hTf formed micelle-like clusters. The molecular modeling confirmed the experimental results. This study is expected to provide important insight into the interaction of hTf with ROP and ASA to use in various toxicological and therapeutic processes.

Investigations with spectroscopy, zeta potential and molecular modeling of the non-cooperative behaviour between cyclophosphamide hydrochloride and aspirin upon interaction with human serum albumin: binary and ternary systems from multi-drug therapy.

The interaction between cyclophosphamide hydrochloride (CYC) and aspirin (ASA) with human serum albumin (HSA) was studied by various kind of spectroscopic, ζ potential and molecular modeling under physiological conditions. The fluorescence data showed that the binding of drugs to proteins caused strong static fluorescence quenching. The analysis of the fluorescence quenching of HSA in the binary and ternary systems displayed that ASA was affected by the complex formed between CYC and HSA. Moreover, CYC was influenced by the HSA-ASA complex. The inherent binding information, including the quenching mechanism, binding constants, number of binding sites, effective quenching constant, fraction of the initial fluorescence and thermodynamic parameters were measured by the fluorescence quenching technique at various temperatures. In addition, according to the synchronous fluorescence spectra of HSA, the results showed that the fluorescence quenching of HSA originated from the Trp and Tyr residues, and indicated a conformational change of HSA with the addition of the drugs. Far-UV CD spectra of HSA were recorded before and after the addition of ASA and CYC as binary and ternary systems. An increase in intensity of the positive CD peak of HSA was observed in the presence of the drugs. The results were interpreted by excited interactions between the aromatic residues of the HSA binding sites and the drugs bound to them. The distance r between donor and acceptor was obtained by the Forster energy according to fluorescence resonance energy transfer (FRET) and found to be 2.35 nm and 1.78 nm for CYC and ASA, respectively. This confirmed the existence of static quenching for proteins in the presence of CYC and ASA. Furthermore, docking studies pointed at a reduction of the affinity of each of the drug compounds to the protein in the presence of the other in meaningful amounts. Pre-binding of any of the said compounds forced the second to bind in a non-optimized location and orientation. The potential at the electrokinetic shear surface of the protein-drug solution were measured at several concentrations of the drugs by the ζ potential technique, which confirmed experimental and theoretical results.

Influence of transmembrane pressure and feed concentration on the retention of arsenic, chromium and cadmium from water by nanofiltration.

One of the main toxic pollutants in drinking water is heavy metals which must be reduced to standard levels. Removal of trace amounts of heavy metals can be achieved by means of membrane processes such as nanofiltration. The removal efficiency of a nanofiltration membrane is strongly affected by operating conditions. The present study focused on the effect of two key parameters, i.e., transmembrane pressure and feed concentration on the removal of heavy metals (arsenic, chromium and cadmium) from water by a polymeric nanofiltration membrane UTC-70UB charged negatively. The rejection experiments included variation of heavy metals feed concentrations in the range of 100 to 400 microg/L for arsenic and chromium and 20 to 80 microg/L for cadmium, and different transmembrane pressures in the range of 5 to 14 bar. The results indicated that under most conditions tested in this research, the rejection of heavy metals was found to increase when the transmembrane pressure was increased. The results also showed the high rejection percentage of the heavy metals, with the maximum retention values of arsenic, cadmium and chromium, 97%, 100% and 95% respectively. The percent reduction of arsenic and chromium was found to enhance as their concentration in the feed increased. However, in the case of cadmium, the rejection was reduced with increase in the concentration.