Kazimierz Gąsiorowski

Chemical structure of phenothiazines and their biological activity.

Phenothiazines belong to the oldest, synthetic antipsychotic drugs, which do not have their precursor in the world of natural compounds. Apart from their fundamental neuroleptic action connected with the dopaminergic receptors blockade, phenothiazine derivatives also exert diverse biological activities, which account for their cancer chemopreventive-effect, as: calmodulin- and protein kinase C inhibitory-actions, anti-proliferative effect, inhibition of P-glycoprotein transport function and reversion of multidrug resistance. According to literature data on relations between chemical structure of phenothiazines and their biological effects, the main directions for further chemical modifications have been established. They are provided and discussed in this review paper.

New dressing materials derived from transgenic flax products to treat long-standing venous ulcers - a pilot study.

A new flax dressing product was developed based on three components (fibers, oil emulsion, and seedcake extract) from genetically engineered flax plants that were obtained by plant transformation using three genes controlling the synthesis of antioxidative compounds from the phenylpropanoid pathway. Simultaneous flax explant transformation with three genes coding for chalcone synthase, chalcone isomerase, and dihydroflavonol reductase resulted in an accumulation of phenolic acids in the fibers, polyunsaturated fatty acids in the oil, and lignans in the seedcake. The fibers, oil, and seedcake from transgenic flax contained a broad spectrum of antioxidative compounds. They were tested for cytotoxicity, and none were found to have a negative effect on the growth and morphology of Balb/3T3 cells. In this preliminary report, we present pilot data on the effects of using linen dressing treatment on its own or in combination with oil emulsion and/or seedcake extract on chronic wound healing. After a 12‐week study, we concluded that an application of a modified flax‐dressing (linen) bandage might yield a more rapid rate of healing and reduce the wound exudes and wound size. In several cases, wound healing was completed during the period of investigation. Interestingly and importantly, the patients reported that the new bandage made from modified flax diminished the pain accompanying chronic venous ulceration. Further study is required to determine any definitive effects of flax bandage on wound healing. This is the first pilot study report suggesting the benefits of a flax‐based dressing on wound healing.

Vascular factors and epigenetic modifications in the pathogenesis of Alzheimer's disease

Alzheimers disease (AD) is a debilitating illness with no known cure. Nowadays accumulating evidence suggested that the vascular endothelium and chronic hypoperfusion may play important role in pathobiology of AD. The vascular endothelium which regulates the passage of macromolecules and circulating cells from blood to tissue, is a major target of oxidative stress, playing a critical role in the pathophysiology of vascular diseases. Since the vascular endothelium, neurons and glia are all able to synthesize, store and release reactive oxygen species (ROS) and vascular active substances in response to certain stimuli, their contribution to the pathophysiology of AD can be very important. New evidence indicates that continuous formation of free ROS induces cellular damage and decreases antioxidant defenses. Specifically, oxidative stress increases vascular endothelial permeability and promotes leukocyte adhesion. We summarize the reports that sporadic, late-onset of AD results from vascular etiology. Recently an involvement of epigenetic alterations in the etiology of AD is also intensively investigated. Gaining a more complete understanding of the essential components and underlying mechanisms involved in epigenetic regulation could lead to novel treatments for a number of neurological and psychiatric conditions.

Phase separation in phosphatidylcholine membrane caused by the presence of a pyrimidine analogue of fluphenazine with high anti-multidrug-resistance activity.

Phenothiazine compounds are known as effective inhibitors of a multidrug resistance (MDR) of tumor cells to chemotherapeutic agents. This group consists of many important substances used in human medicine such as antipsychotic drugs in the case of fluphenazine (FPh) or chlorpromazine (CPZ). Fluphenazine was on the World Health Organization (WHO) list of Essential Medicines of 2009, and its new pyrimidine analog (FPh-prm) presented in this work has been documented to have a high anti-MDR activity. In order to discover the character of alterations of the lipid bilayer structure caused by the presence of FPh-prm inside the lipid membrane, which is responsible for the essential increase of an anti-MDR activity of FPh-prm, microcalorimetric (differential scanning calorimetry), Laurdan fluorescence, (31)P nuclear magnetic resonance spectroscopy (NMR), and attenuated total reflectance Fourier transfer infrared spectroscopy (FTIR-ATR) were used for dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) liposomes mixed with a different concentration of amine analogue. It was stated that the formation of domains with different content of FPh-prm/DPPC can be a reason for the membrane-related mechanism of chemoprevention associated with the inhibition of the outward transport of anticancer drugs by the glycoprotein P (Pgp) in cancer cells by the pyrimidine analog of FPh. To our best knowledge, this report is the first to show the bilayer structure of domains formed by incomplete miscibility of fluphenazine-related compounds and phospholipid molecules. Our results provide a sound basis for the design of future modifications of anti-MDR drugs by providing very effective inhibitors of the pump activity of Pgp.

RAGE-TLR Crosstalk Sustains Chronic Inflammation in Neurodegeneration

Chronic inflammatory reactions are consistenly present in neurodegeneration of Alzheimer type and are considered important factors that accelerate progression of the disease. Receptors of innate immunity participate in triggering and driving inflammatory reactions. For example, Toll-like receptors (TLRs) and receptor for advanced glycation end product (RAGE), major receptors of innate immunity, play a central role in perpetuation of inflammation. RAGE activation should be perceived as a primary mechanism which determines self-perpetuated chronic inflammation, and RAGE cooperation with TLRs amplifies inflammatory signaling. In this review, we highlight and discuss that RAGE-TLR crosstalk emerges as an important driving force of chronic inflammation in Alzheimer’s disease.

Immunomodulating polysaccharide fractions of Menyanthes trifoliata L.

Abstract Looking for new plant sources of immunomodulating agents polysaccharide-rich fractions (PS) from Menyanthes trifoliata L. (Menyanthaceae) have been isolated. The herb of Menyanthes trifoliata L. was sequentially extracted with water, 0.1 м NaOH, 8% CH3COOH, and 1 м NaOH. After dialysis and resolution on Biogel P-10 four homogenic (B-4, B-5, C-4, D-5) and two nonhomogenic (A-3 and D-4) PS were isolated. About 0.5% of PS over 3500 Da were found in the dry plant material. They were characterized through chemical analysis, NMR and vibrational spectroscopy. Speciation analysis of chosen metal/metaloid elements was performed and an exceptionally high concentration of Se was found in PS of a pure water extract (A-3). The biological tests on the immunomodulating influence with human blood-derived lymphocytes and granulocytes revealed that two fractions, B-4 and B-5, were strong stimulators of immune cells, whereas fractions D-5 and A-3 were found as potent suppressive and anti-inflammatory agents. The applied isolation procedures led to the separation of active compounds into stimulatory and inhibitory fractions.

New fluphenazine analogues as inhibitors of P-glycoprotein in human lymphocyte cultures

Aim of the study To evaluate the inhibitory effect of 17 new analogues of FPh on the Pgp transport function, by estimation of the rhodamine 123 (Rod-123) accumulation inside cultured lymphocytes. Material and methods Lymphocyte were cultured in the presence of a lectin (PHA; 2%, v/v), incubated with benzo[α]pyrene (B[α]P; 7.5 µM, 48 h) to induce genotoxic damage and to increase Pgp expression in the cells. Lymphocytes cultured without the tested compounds were considered as controls. Results It was established that 10 analogues of FPh, among 17 tested, significantly increased Rod-123 accumulation in lymphocytes at the concentration of 10 µM. As compared to the control cultures the Pgp transport function was the most strongly inhibited by 1a, 1b, 1d, 3f, 3h and 3i analogues (approximately by 25%). Conclusions FPh analogues 1a, 1b, 1d, 3f, 3h and 3i should be further studied as promising candidates for adjuvant cancer chemotherapeutics.

Synthesis, COX-1/2 inhibition activities and molecular docking study of isothiazolopyridine derivatives

One of the main challenges for nowadays medicine is drugs selectivity. In COX-1 and COX-2, the active sites are composed of the same group of amino acids with the exception of the only one residue in position 523, in COX-1 is an isoleucine, while in COX-2 is a valine. Here, we presented a series of isothiazolopyridine/benzisothiazole derivatives substituted differently into an isothiazole ring, which were synthesized and investigated for their potencies to inhibit COX-1 and COX-2 enzymes by colorimetric inhibitor screening assay. All the tested compounds inhibited the activity of COX-1, the effect on COX-2 activity was differential. The mode of binding was characterized by a molecular docking study. Comparing biological activity of the investigated compounds, it was observed that compounds sharing the most similar position to flurbiprofen and meloxicam, representing the two main enzyme subdomains, achieved higher biological activity than others. It is directly related to the fit to the enzymes active site, which prevents too early dissociation of the compounds.

Emulsions Made of Oils from Seeds of GM Flax Protect V79 Cells against Oxidative Stress

Polyunsaturated fatty acids, sterols, and hydrophilic phenolic compounds are components of flax oil that act as antioxidants. We investigated the impact of flax oil from transgenic flax in the form of emulsions on stressed Chinese hamster pulmonary fibroblasts. We found that the emulsions protect V79 cells against the H2O2 and the effect is dose dependent. They reduced the level of intracellular reactive oxygen species and protected genomic DNA against damage. The rate of cell proliferation increased upon treatment with the emulsions at a low concentration, while at a high concentration it decreased significantly, accompanied by increased frequency of apoptotic cell death. Expression analysis of selected genes revealed the upregulatory impact of the emulsions on the histones, acetylases, and deacetylases. Expression of apoptotic, proinflammatory, and anti-inflammatory genes was also altered. It is thus suggested that flax oil emulsions might be useful as a basis for biomedical products that actively protect cells against inflammation and degeneration. The beneficial effect on fibroblast resistance to oxidative damage was superior in the emulsion made of oil from transgenic plants which was correlated with the quantity of antioxidants and squalene. The emulsions from transgenic flax are promising candidates for skin protection against oxidative damage.

Fluphenazine: from an isolated molecule to its interaction with lipid bilayers

Fluphenazine (FPh) belongs to the phenothiazine family of compounds and exhibits a wide variety of biological effects, including antimutagenic, proapoptotic, antiproliferative and anti-multidrug resistance (MDR) activities. The ability of FPh to interact with lipid membranes can have a significant impact on its biological activities. However, the mechanisms involved in the interaction of FPh with lipid membranes are poorly understood. FTIR-ATR spectroscopy has been used in this study to visualize the interactions between FPh and a model lipid bilayer composed of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). Subsequent interpretation of the temperature-dependent FTIR spectra obtained for FPh:DPPC systems containing different concentrations of FPh was efficiently supported by principal component analysis.