Synthesis,receptor binding studies,optical spectroscopic and in silico structural characterization of morphiceptin analogs with cis‐4‐amino‐L‐proline residues

Three novel morphiceptin analogs, in which Pro in position 2 and/or 4 was replaced by cis‐4‐aminoproline connected with the preceding amino acid through the primary amino group, were synthesized. The opioid receptor affinities, functional assay results, enzymatic degradation studies and experimental and in silico structural analysis of such analogs are presented. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.     

Transformation of Curvularia lunata IM 2901 with pAN7-1 influences selected physiological properties of the fungus

Genetic analysis of Curvularia lunata IM 2901 transformants, previously obtained by electroporation with plasmid pAN7-1, was carried out. Isolates displayed several differences in hygromycin B resistance and their physiology. It was shown that plasmid pAN7-1 was integrated in different copy numbers and at different positions in the genome of the strains studied. Both the wild type and pAN7-1 isolates, when growing in liquid media, produced an extracellular emulsifying agent. The transformants differed in their growth kinetics, intensity of surfactant production and in the efficiency of cortexolone 11beta-hydroxylation, in comparison with the wild type. The micro-organisms varied in susceptibility to the lytic enzyme complex (Novozyme 234), which indicated the presence of differences in their cell wall composition and/or in architecture caused by an integrated plasmid pAN7-1.     

Microbial analysis of clinical material taken from patients at the Oncology Center, Maria Skłodowkda-Curie Institute in Warsaw in 1997

An analysis was carried out of the microbiological investigations of clinical material samples obtained from the patients of two oncology centres belonging to the Warsaw Oncology Centre. Microorganisms cultured from urine, blood, catheters, smears of wounds and other materials were analysed. From 4839 clinical material samples from the Ursynów centre 1755 bacterial strains were isolated. From 423 samples from the centre in Wawelska Street 171 strains were obtained. In infections of patients from the centres the number of Gram-positive cocci was twice that of Gram-negative rods. In the investigated clinical material S. aureus was the most frequently isolated Gram-positive coccus, while E. coli was the most frequent species among Gram-negative bacteria. In the infections of oncological patients a considerable frequency was noted of yeast-like fungi, especially C. albicans. Particularly disquieting was the increasing number of isolates of C. glabrata and C. krusei strains resistant to fluconazole.     

Adeno-associated viruses (AAV)

Recombinant adeno-associated virus derived vectors (rAAV) a thought to be a most promising candidates for gene therapy applications. Their nonpathogenic nature as well as the encouraging capability to infect both proliferating and non proliferating cells are advantages for gene therapy applications. Here, we summarize the potential mechanisms responsible for AAV maintenance and site-specific integration to human genome. The role of Rep proteins, inverted terminal repeats and p5 promotor sequences for chromosomal incorporation of AAV are discussed. Making the site-specific integrative recombinant AAV vectors for gene therapy seems to be closely dependent on the development of viral vectorology.     

Nested PCR for the detection of Aspergillus species in maxillary sinus samples of patients with chronic sinusitis

Background

Fungal rhinosinusitis has become an increasingly recognized disease, being Aspergillus species responsible for most of the cases. Its diagnosis is quite difficult because of the non-specific symptoms and low sensitivity of the current diagnostic methods.

The effect of new non-cross resistant antitumour agents on the energy state of human erythrocytes

Multidrug resistance (MDR) of tumour cells is related to the overexpression of ATP-dependent pumps responsible for the active efflux of antitumour agents out of resistant cells. Benzoperimidine and anthrapyridone compounds exhibit comparable cytotoxic activity against sensitive and MDR tumour cells. They diffuse extremely rapidly across the plasma membrane and render the ATP-dependent efflux inefficient. Such uptake could disturb an energy metabolism of normal cells possessing an elevated level of ATP-dependent proteins, especially erythrocytes having a high level of the MRP1, MRP4 and MRP5 proteins. In this study the effect of five antitumour agents: benzoperimidine (BP1), anthrapyridones (CO1, CO7) and reference drugs used in the clinic: doxorubicin (DOX) and pirarubicin (PIRA), on the energetic state in human erythrocytes has been examined. These compounds have various types of structure and kinetics of cellular uptake (slow--DOX, CO7, moderate--PIRA, fast--BP1, CO1) resulting in their different ability to saturate ATP-dependent transporters. The energetic state of erythrocytes was examined by determination of purine nucleotide contents (ATP, ADP, AMP), NAD(+) and values of adenylate energy charge (AEC) using an HPLC method. It was found that the level of nucleotides as well as the AEC value of erythrocytes were not changed during 24 h of incubation with these agents independently of their structure and ability to saturate ATP-dependent pumps. This is a very promising result in view of their potential use in the clinic as antitumour drugs against multidrug resistant cancers.     

Purification of antibodies against N-homocysteinylated proteins by affinity chromatography on Nomega-homocysteinyl-aminohexyl-Agarose

Modification with homocysteine (Hcy)-thiolactone leads to the formation of N-Hcy-Lys-protein. Although N-Hcy-Lys-proteins are immunogenic, pure antibodies have not yet been obtained. Here we describe synthesis and application of Nomega-homocysteinyl-aminohexyl-Agarose for affinity purification of anti-N-Hcy-Lys-protein antibodies. Nomega-homocysteinyl-aminohexyl-Agarose was prepared by N-homocysteinylation of omega-aminohexyl-Agarose with Hcy-thiolactone. Immune serum was obtained from rabbits inoculated with N-Hcy-Lys-keyhole limpet hemocyanine and IgG fraction prepared by chromatography on protein A-Agarose. Anti-N-Hcy-Lys-protein IgG was adsorbed on Nomega-homocysteinyl-aminohexyl-Agarose column at pH 8.6 and eluted with a pH 2.3 buffer. Enzyme-linked immunosorbent assays demonstrate that the antibody recognizes specifically N-homocysteinylated variants of hemoglobin, albumin, transferrin, and antitrypsin.     

Biotransformation of 4‐fluoro‐N‐(1‐{2‐[(propan‐2‐yl)phenoxy]ethyl}‐8‐azabicyclo[3.2.1]octan‐3‐yl)‐benzenesulfonamide,a novel potent 5‐HT7 receptor antagonist with antidepressant‐like and anxiolytic properties: In vitro and in silico approach

The aim of the study was to investigate the metabolism of 4‐fluoro‐N‐(1‐{2‐[(propan‐2‐yl)phenoxy]ethyl}‐8‐azabicyclo[3.2.1]octan‐3‐yl)‐benzenesulfonamide (PZ‐1150), a novel 5‐HT₇ receptor antagonist with antidepressant‐like and anxiolytic properties, by the following three ways: in vitro with microsomes; in vitro employing Cunninghamella echinulata, and in silico using MetaSite. Biotransformation of PZ‐1150 with microsomes resulted in five metabolites, while transformation with C. echinulata afforded two metabolites. In both models, the predominant metabolite occurred due to hydroxylation of benzene ring. In silico data coincide with in vitro experiments, as three MetaSite metabolites matched compounds identified in microsomal samples. In human liver microsomes PZ‐1150 exhibited in vitro half‐life of 64 min, with microsomal intrinsic clearance of 54.1 μL/min/mg and intrinsic clearance of 48.7 mL/min/kg. Therefore, PZ‐1150 is predicted to be a high‐clearance agent. The study demonstrated the applicability of using microsomal model coupled with microbial model to elucidate the metabolic pathways of compounds and comparison with in silico metabolite predictions.