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Oxidative stress and inflammation are DNA instability factors for rheumatoid arthritis (RA) patients. The aims of this study were to evaluate cytogenetic alterations in Peripheral Blood Lymphocytes (PBL) in two groups of RA patients: the early and the long-term RA group; and to examine potential of concomitant treatment with Methotrexate (MTX) and Dry olive leaf extract (DOLE) against cytogenetic damage in RA patients after a 3-weeks treatment. A total of 32 RA patients and 10healthy individuals were included. RA patients were equally divided into four groups: two groups with early phase RA (one treated with MTX alone, the other in combination with DOLE); and two long-term phase RA groups (group with active disease and group with low disease activity)-both treated with MTX and DOLE combination. PBL cultures were screened for chromosome aberrations and micronuclei frequencies. Significantly increased frequencies of micronuclei were shown in active phase RA disease (both early and long-term) but not in the group with low disease activity, as compared to controls. Chromosome aberrations were detected for all 4 RA groups. The highest frequencies of micronuclei and chromosome aberrations were found in the long-termactive RA group. After 3 weeks-treatment, there were no significant decrease of the micronuclei frequencies compared to baseline, although they were reduced in all RA groups, except for the group with the long-term active disease. High level of cytogenetic damage in RA patients was concordant with duration and activity of the RA disease. At 3 weeks of therapy, neither the combined treatment (MTX+DOLE), nor MTX alone did not affect the frequency of micronuclei formation. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. OI 173034]

More than 30 types of amyloids are linked to close to 50 diseases in humans, the most prominent being Alzheimer’s disease (AD). AD is brain-related local amyloidosis, while another amyloidosis, such as AA amyloidosis, tends to be more systemic. Therefore, we need to know more about the biological entities’ influencing these amyloidosis processes. However, there is currently no support system developed specifically to handle this extraordinarily complex and demanding task. To acquire a systematic view of amyloidosis and how this may be relevant to the brain and other organs, we needed a means to explore "amyloid network systems" that may underly processes that leads to an amyloid-related disease. In this regard, we developed the DES-Amyloidoses knowledgebase (KB) to obtain fast and relevant information regarding the biological network related to amyloid proteins/peptides and amyloid-related diseases. This KB contains information obtained through text and data mining of available scientific literature and other public repositories. The information compiled into the DES-Amyloidoses system based on 19 topic-specific dictionaries resulted in 796,409 associations between terms from these dictionaries. Users can explore this information through various options, including enriched concepts, enriched pairs, and semantic similarity. We show the usefulness of the KB using an example focused on inflammasome-amyloid associations. To our knowledge, this is the only KB dedicated to human amyloid-related diseases derived primarily through literature text mining and complemented by data mining that provides a novel way of exploring information relevant to amyloidoses.

Oxidative damage to protein and lipid macromolecules in target organs in hypertension has been recognized as a major factor contributing to cardiovascular, cerebrovascular, and renal diseases. Data on protein and lipid oxidative damage in spontaneously hypertensive rats are numerous, but there is no information on DNA damage in tissues measured by comet assay. The aim of this study was to determine the baseline damage to DNA, protein, and lipid macromolecules in different organs of spontaneously hypertensive rats. Markers of lipid peroxidation, protein oxidation, and DNA damage were measured in blood, heart, kidney, and liver of 24-week-old spontaneously hypertensive rats. Plasma prooxidant and antioxidant status were determined as well. Age-matched normotensive Wistar rats were used as control. A rise in markers of lipid peroxidation and protein oxidation, malondialdehyde, and advanced oxidation protein products, was detected in all tissues of spontaneously hypertensive rats, with particularly high values in the liver. DNA damage, measured by the comet assay, was significantly higher in all the studied tissues of spontaneously hypertensive rats compared to normotensive control, with more severe damage in the cardiac and renal cells. Significant depletion of the plasma antioxidant barrier in spontaneously hypertensive rats was also observed. This study showed increased damage to all macromolecules in all studied samples of spontaneously hypertensive rats in comparison with control Wistar rats.

Ganoderma lucidum is traditionally used in Eastern medicine to preserve vitality, promote longevity, and treat disease. It possesses immunomodulatory, antitumor, antimicrobial, and antiaging activities, among others, but one of the most important is its antioxidant property, which is the basis for other effects, because free radicals trigger many diseases. The substrate commonly used for commercial cultivation of G. lucidum is not environmentally friendly nor economically justified, so there is a need to find new alternative substrates. The aim of this study was to analyze the effect of substrate composition on the bioactivity of G. lucidum basidiocarps. G. lucidum was cultivated on 2 different substrates: (1) a mixture of wheat straw, grapevine branches, and wheat bran, and (2) wheat straw. Commercial fruiting bodies, cultivated on oak sawdust, were used as the control. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, total phenols, and flavonoid content were determined spectrophotometrically to define the antioxidative potential of basidiocarp extracts. The comet test was performed to detect the degree of DNA damage in the cells that were exposed to G. lucidum extracts before and after the effect of oxidants. Higher antioxidative potential was observed for the extract of G. lucidum basidiocarps cultivated on wheat straw compared with that from the mixed substrate and especially with commercial ones. The alternatively cultivated basidiocarps also showed stronger antigenotoxic potential compared with commercial ones. The study showed that fruiting bodies produced on wheat straw, one of the most accessible and cheapest crop residues, are more potent antioxidant and antigenotoxic agents than commercially cultivated ones.

To remedy carotid artery stenosis and prevent stroke surgical intervention is commonly used, and the gold standard being carotid endarterectomy (CEA). During CEA cerebrovascular hemoglobin oxygen saturation decreases and when this decrease reaches critical levels it leads to cerebral hypoxia that causes neuronal damage. One of the proposed mechanism that affects changes during CEA and contribute to acute brain ischemia (ABI) is oxidative stress. The increased production of reactive oxygen species and reactive nitrogen species during ABI may cause an unregulated inflammatory response and further lead to structural and functional injury of neurons. Antioxidant activity are involved in the protection against neuronal damage after cerebral ischemia. We hypothesized that neuronal injury and poor outcomes in patients undergoing CEA may be results of oxidative stress that disturbed function of antioxidant enzymes and contributed to the DNA damage in lymphocytes. © 2019 The Authors

The objective of this work was to investigate and profoundly characterize low-energy nanoemulsions as multifunctional carriers, with slight reference to dermal administration. An evidence-based approach was offered for deepening the knowledge on their formation via spontaneous emulsification. Curcumin, a compound of natural origin, potentially powerful therapeutic, was chosen as a model API. Due to curcumin's demanding properties (instability, poor solubility, low permeability), its potentials remain unreached. Low-energy nanoemulsions were considered carriers capable of overcoming imposed obstacles. Formulation consisting of Polysorbate 80 and soybean lecithin as stabilizers (9:1, 10%), medium-chain triglycerides as the oil phase (10%) and ultrapure water was selected for curcumin incorporation in 3 different concentrations (1, 2 and 3 mg/mL). Physicochemical stability was demonstrated during 3 months of monitoring (mean droplet size: 111.3-146.8 nm; PDI LT 0.2; pH: 4.73-5.73). Curcumin's release from developed vehicles followed Higuchi's kinetics. DPPH (IC50 = 0.1187 mg/ mL) and FRAP (1.19 +/- 0.02 mmol/g) assays confirmed that curcumin acts as a potent antioxidant through different mechanisms, with no alterations after incorporation in the formulation. High biocompatibility in line with antigenotoxic activity of curcumin-loaded formulations (protective and reparative) was estimated through Comet assay. A multidisciplinary approach is needed to fully characterize developed systems, directing them to more concrete application possibilities. This is the peer-reviewed version of the article: I. Nikolic, D. Jasmin Lunter, D. Randjelovic, et. al., Curcumin-loaded low-energy nanoemulsions as a prototype of multifunctional vehicles for different administration routes: physicochemical and in vitro peculiarities important for dermal application, International, Journal of Pharmaceutics, 2018, 550, 1-2, 333-346, DOI: [https://doi.org/10.1016/j.ijpharm.2018.08.060] The published version: [https://cer.ihtm.bg.ac.rs/handle/123456789/2425]

Enhanced vascularization at sensor interfaces can improve long-term function. Fibrin, a natural polymer, has shown promise as a biomaterial for sensor coating due to its ability to sustain endothelial cell growth and promote local vascularization. However, the culture of cells, particularly endothelial cells (EC), within 3D scaffolds for more than a few days is challenging due to rapid loss of EC viability. In this manuscript, a robust method for developing fibrin microbead scaffolds for long-term culture of encapsulated ECs is described. Fibrin microbeads are formed using sodium alginate as a structural template. The size, swelling and structural properties of the microbeads were varied with needle gauge and composition and concentration of the pre-gel solution. Endothelial colony-forming cells (ECFCs) were suspended in the fibrin beads and cultured within a perfusion bioreactor system. The perfusion bioreactor enhanced ECFCs viability and genome stability in fibrin beads relative to static culture. Perfusion bioreactors enable 3D culture of ECs within fibrin beads for potential application as a sensor coating.

Poly (lactic-co-glycolic acid) (PLGA)-based materials are widely investigated for drug delivery and tissue engineering applications. Despite their popularity the genotoxic potential of PLGA has not been investigated. In this study, the comet assay, a sensitive assay for DNA damage, was used to evaluate potential genotoxicity in model cell types exposed to PLGA microspheres. Human umbilical vein endothelial cells (HUVECs) and mesenchymal stem cells (MSCs) cells were exposed to PLGA microspheres (0.4-6 mg/mL) and DNA damage assessed at 24 h, 4 days, and 7 days. DNA damage was not identified after 24 h. However, after 4 and 7 days of exposure to 2 and 6 mg/mL of PLGA microspheres a significant elevation of DNA damage in both cell types was observed. The PLGA microspheres did not exhibit any cytotoxic effects on the cells under the conditions tested. Our results suggest that PLGA may have a genotoxic effect on cells. A broader investigation of the PLGA genotoxic profile in biological systems is needed. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 284-291, 2017.

While Alzheimer’s disease (AD) is primarily considered to be a neurodegenerative disorder, there is much evidence supporting the idea that it is also a systematic disease, so the occurrence of genomic instability in cells that are not neurons is equally important. Accordingly, acrocentric chromosomes play an important role in the etiology of AD, especially the presence of trisomy 21. The aim of this work was to assess the relationship between the frequency of acrocentrics included in satellite association (SA) and acrocentrics with premature centromere division (PCD) in the peripheral blood lymphocytes of AD patients and age-matched controls. Since our results showed that there are significant differences in SA frequency, as well as in the frequency of PCD in acrocentrics between AD patients and control group, we may conclude that the occurrence of acrocentric chromosome alterations presented in this study could be related to the etiology of AD. [Projekat Ministarstva nauke Republike Srbije, br. OI 173034] This article has been corrected. Link to the correction 10.2298/ABS150713084E

Author thanks the Fulbright Association , German DAAD Association and the Ministry of Education, Science and Technological Development of the Republic of Serbia for financial support (Project No: 173034).

Alzheimer?s disease (AD), as the most common form of dementia, has for many years attracted the attention of researchers around the world, primarily because of the problems of reliable diagnostic methods that could help in the early detection of this devastating disease. One of the important aspects of genetic research related to AD is the analysis of chromosome instability which includes: aneuploidies of different chromosomes, telomere shortening and the phenomenon of premature centromere division (PCD). The aim of this study was to describe specific biomarkers in different types of cells as potential parameters for the diagnosis of AD in order to promptly recognize pre-symptomatic stages and prevent the development of disease and/or slow down its progression.

Thyroid hormones stimulate aerobic metabolism which may lead to oxidative stress accompanied by damage to various cellular macromolecules, including DNA. Previous comet assay studies have shown that thyroid hormones cause DNA damage due to the creation of reactive oxygen species (ROS). However, cytogenetic studies have been equivocal because although an increase in the sister-chromatid exchange frequency per cell has been reported increased micronuclei frequency has not. We used cytogenetic examination of chromosome breakage and aberrations in whole-blood cultures of human peripheral blood lymphocytes to investigate possible clastogenic effects when lymphocytes were exposed to 0.002 mu M to 50 mu M of L-thyroxine for 24 h and 48 h, these concentrations being chosen because they had been used in previous studies of sister-chromatid exchange and micronuclei frequency. Under our experimental conditions thyroxine did not induced any statistically significant increase in chromosome breakage or aberrations. This lack of clastogenic effects is in contrast to the reported comet assay results obtained using purified lymphocytes, possibly because whole-blood cultures contain catalase and glutathione peroxidase capable of reducing the effects of reactive oxygen species.

Hormones are cellular products involved in the regulation of a large number of processes in living systems, and which by their actions affect the growth, function and metabo­lism of cells. Considering that hormones are compounds normally present in the organism, it is important to determine if they can, under certain circumstances, lead to genetic changes in the hereditary material. Numerous experimental studies in vitro and in vivo in different systems, from bacteria to mammals, dealt with the mutagenic and genotoxic effects of hormones. This work presents an overview of the research on genotoxic effects of non­steroidal hormones, although possible changes of genetic material under their influence have not still been known enough, and moreover, investigations on their genotoxic influ­ence have given conflicting results. The study results show that mechanisms of genotoxic effect of nonsteroidal hormones are manifested through the increase of oxidative stress by arising reactive oxygen species. A common mechanism of ROS occurence in thyroid hormones and catecholamines is through metabolic oxidation of their phenolic groups. Mani­festation of insulin genotoxic effect is based on production of ROS by activation of NADPH isophorms, while testing oxytocin showed absence of genotoxic effect. Considering that the investigations on genotoxicity of nonsteroidal hormones demonstrated both positive and negative results, the explanation of this discordance involve limitations of test systems themselves, different cell types or biological species used in the experiments, different lev­el of reactivity in vitro and in vivo, as well as possible variations in a tissue-specific expres­sion. Integrated, the provided data contribute to better understanding of genotoxic effect of nonsteroidal hormones and point out to the role and mode of action of these hormones in the process of occurring of effects caused by oxidative stress. Hormoni su ćelijski proizvodi uključeni u regulaciju velikog broja procesa u živim sistemima koji svojim dejstvom utiču na rast, funkciju ili metabolizam ćelija. Obzirom da su hormoni jedinjenja koja su uobičajeno prisutna u organizmu, značajno je utvrditi da li oni mogu pod izvesnim okolnostima dovesti do genetičkih promena na naslednom materijalu. Eksperimentalna ispitivanja u in vitro i in vivo uslovima u različitim sistemima, od bakterija do sisara, bavila su se istraživanjem mutagenih i genotoksičnih efekata hormona. U ovom radu je dat pregled istraživanja genotoksičnih efekata nesteroidnih hormona, pošto još uvek nisu dovoljno poznate moguće promene naslednog materijala pod njihovim uticajem, a i ispitivanja njihovog genotoksičnog dejstva su dala oprečne rezultate. Rezultati studija pokazuju da se mehanizmi genotoksičnog dejstva nesteroidnih hormona ispoljavaju kroz povećanje oksidativnog stresa nastankom reaktivnih vrsta kiseonika (reactive oxygen species - ROS). Uobičajeni mehanizam nastanka ROS kod tireoidnih hormona i kateholamina je putem metaboličke oksidacije njihovih fenolnih grupa. Ispoljavanje genotoksičnog efekta insulina se zasniva na produkciji ROS putem aktivacije NADPH izoformi, dok je ispitivanje oksitocina pokazalo odsustvo genotoksičnog efekta. Uzimajući u obzir da su ispitivanja genotoksičnosti nesteroidnih hormona pokazala i pozitivne i negativne rezultate, objašnjenja ovog neslaganja obuhvataju ograničenja samih test sistema, različite tipove ćelije ili bioloških vrsta upotrebljenih u eksperimentima, različiti nivo reaktivnosti u in vitro i in vivo uslovima, kao i moguće razlike u tkivno specifičnoj ekspresiji. Objedinjeni, navedeni podaci doprinose boljem razumevanju genotoksičnih efekata nesteroidnih hormona i ukazuju na ulogu i načine delovanja ovih hormona u procesu nastanka efekata izazvanih oksidativnim stresom.

Aging is the main risk factor for Alzheimer’s disease (AD). With aging, inflammation has been recognized as potential trigger for starting the neurodegenerative cascade leading to neuronal death. Before Aβ and tau accumulation, evidence has put alterations of the cell cycle at the core of these processes. Still, a number of features of the cell cycle re-entry phenotype have remained elusive to the role of ectopic protein expression in the process of neuroinflammation and consequently neuronal cell death. Recently, a novel cyclin dependent kinase CDK11 has been found to be involved in astrocyte mediated inflammatory response and Alzheimer’s disease. In this review, we aim to establish the missing part of the puzzle between neuroinflammation and APP / Aβ deregulation in AD by evaluating the role of a cyclin, CDK11. CDK11 may play a vital role in cell cycle re-entry in AD neurons in an APP-dependent manner, thus presenting an intriguing novel function of the APP signaling pathway in AD.

X-chromosome instability has been a long established feature in Alzheimer's disease (AD). Premature centromere division and aneuploidy of the X-chromosome has been found in peripheral blood lymphocytes and neuronal tissue in female AD patients. Interestingly, only one chromosome of the X pair has been affected. These results raised a question, "Is the X-chromosome inactivation pattern altered in peripheral blood lymphocytes of women affected by AD?" To address this question, we analyzed the methylation status of androgen receptor promoter which may show us any deviation from the 50 : 50% X inactivation status in peripheral blood lymphocytes of women with AD. Our results showed skewed inactivation patterns (>90%). These findings suggest that an epigenetic alteration on the inactivation centers of the X-chromosome (or skewing) relates not only to aging, by might be a novel property that could account for the higher incidence of AD in women.

Chromosomal alterations as a sign of genetic instability are a feature of Alzheimer's disease (AD). Assessment of the genetic instability of non-neuronal cells of AD patients may provide a method to diagnose or monitor prognosis of the disease. Considering the importance of X chromosome alterations in the possible etiology of AD females, we used fluorescent in situ hybridization (FISH) for the centromere region of the X chromosome to determine aneuploidy, for a possible correlation with premature centromere division (PCD, X) in lymphocytes of AD females and age-matched controls. In AD patients, our results showed a marked and significant increase in the frequency of the X chromosome aneuploidy comparing with age matched controls (p lt 0.001). Also, a significant difference was detected in the PCD, X frequency between AD females when compared with age matched controls (p lt 0.001). In addition, a strong (R2=0.97, n=20) and significant (p lt 0.001) correlation was found between the frequency of aneuploidy and PCD, X in the AD group. Our results support the view that AD is a generalized systematic disease where PCD is to be considered as a stable sign of disease leading to aneuploidy.

Cytogenetic analysis of the X chromosome in phytohaemagglutinin stimulated peripheral blood lymphocytes was evaluated in 12 sporadic Alzheimer disease (AD) patients and in 11 healthy subjects. For chromosome analysis two methods were used: (1) standard analysis of G-banded metaphase chromosomes and; (2) fluorescent in situ hybridization (FISH) for the detection of the X chromosome centromeric region in interphase nuclei. Cytogenetic analysis revealed that the X chromosome expresses premature centromere division (PCD) in AD females in 10.53% of metaphase cells and in 15.22% of interphase nuclei. In AD men the percentages were 3.98 and 6.06%, respectively. X chromosome PCD in the female control group showed a percentage of 7.46% in metaphase cells and 9.35% in interphase nuclei and in male controls the percentages were 2.84% in metaphases and 5.54% in interphase nuclei. The results of FISH analysis showed that PCD could occur much earlier than metaphase of mitosis, i.e. in interphase of the cell cycle, immediately after replication. The FISH method can be used for PCD verification in all phases of the cell cycle in various disorders including AD.

Years ago, it was thought that a genetic component was the fundamental cause of a number retinopathy diseases including age related macular degeneration (AMD). Since then, information has emerged about novel genes that contribute to various forms of AMD and other retinopathies that have been eluding researchers for years. In the genetic sense, only the APOE 2 and 4 genes have been found to be a risk factor for sporadic AMD. But, a recent Genome wide association study (GWAS) revealed that an alteration of five SNIPs on the X chromosome in a gene named DIAPH2 may be a susceptibility gene for AMD. Furthermore, the gene DIAPH2 showed to have a polygenic pleiotropy for premature ovarian failure (POF) and AMD in a cohort of women. POF is highly associated with X chromosome skewing, an epigenetic alteration of the inactivation process of the X chromosome. These findings suggest a hypothesis that an epigenetic alteration on the inactivation centres of the X chromosome (or skewing) relates not only to aging, but might be a novel property that affects women with AMD more often than men.

Alzheimer’s disease (AD) is the most frequent progressive neurodegenerative disorder in elderly associated with irreversible cognitive impairment and dementia. The vast majority of AD patients are sporadic (SAD) in which the disease develops after age of 65. Despite of century of research, we lack understanding of the SAD etiology and pathogenesis. Several hypotheses try to explain the main causes of brain degeneration in SAD, one of them assuming that genomic instability and the reentry of certain neurons into the incomplete cell cycle may be the pathogenic basis of the disease. Although the brain is the most affected organ in AD, numerous studies showed structural and functional alterations in peripheral tissues, suggesting that AD is a generalized systemic disorder. Diverse changes in peripheral cells from AD patients are described in literature including cell cycle aberration and chromosome instability, alterations in cell viability, proliferation and apoptosis, oxidative metabolism, amyloid precursor protein and amyloid β protein metabolism, and other cellular processes. The aim of this paper was to summarize and review the results of our investigations and the growing literature data concerning the multiple chromosomal alterations in peripheral cells of AD patients and to consider their possible role in the disease pathogenesis as well as the importance of such investigations. [Projekat Ministarstva nauke Republike Srbije, br. 173034]

Excessive release of stress hormone adrenaline is accompanied by generation of reactive oxygen species which may cause disruption of DNA integrity leading to cancer and age-related disorders. Phenolic-rich plant product dry olive leaf extract (DOLE) is known to modulate effects of various oxidants in human cells. The aim was to evaluate the effect of commercial DOLE against adrenaline induced DNA damage in human leukocytes by using comet assay. Peripheral blood leukocytes from 6 healthy subjects were treated in vitro with three final concentrations of DOLE (0.125, 0.5, and 1 mg/mL) for 30 min at 37 degrees C under two different protocols, pretreatment and post-treatment. Protective effect of DOLE was assessed from its ability to attenuate formation of DNA lesions induced by adrenaline. Compared to cells exposed only to adrenaline, DOLE displayed significant reduction (P lt 0.001) of DNA damage at all three concentrations and under both experimental protocols. Pearson correlation analysis revealed a significant positive association between DOLE concentration and leukocytes DNA damage (P lt 0.05). Antigenotoxic effect of the extract was more pronounced at smaller concentrations. Post-treatment with 0.125 mg/mL DOLE was the most effective against adrenaline genotoxicity. Results indicate genoprotective and antioxidant properties in dry olive leaf extract, strongly supporting further explorations of its underlying mechanisms of action.

Ephedrine, a natural alkaloid from plants of the genus Ephedra, has a chemical structure similar to catecholamines. It is well established that catecholamines (adraneline, noradrenaline and dopamine) cause genotoxic and mutagenic effects. Therefore, the objectives of this investigation were to examine weather ephedrine can exhibit genotoxic effects on isolated human lymphocytes in the Comet assay. Dose-response of human lymphocytes was determined at the concentration range of ephedrine from 0.0005 μM to 500 μM. Three concentrations of ephedrine (1, 50 and 300 μM) which had acceptable cell viability (over 90%) were used for further experiments with inhibitors of DNA reparation (cytosine arabinoside and hydroxyurea). The obtained results showed that ephedrine did not induce DNA damage in isolated human lymphocytes. However, co-treatment of the negative control with DNA repair inhibitors caused a slight but significant increase of DNA damage, due to an endogenous DNA damage. Interestingly, cells treated with ephedrine and DNA repair inhibitors did not express increased DNA damage. On the basis of the obtained results it can be concluded that ephedrine did not exhibit genotoxic effects on isolated human lymphocytes. This result is in accordance with previous investigations showing negative genotoxicological results for ephedrine using bacterial gene mutation test-systems and in vitro cytogenetic analysis. Efedrin, prirodni alkaloid iz biljaka roda Ephedra, ima sličnu hemijsku strukturu sa kateholaminima. Dobro je poznato da kateholamini (adrenalin, noradrenalin i dopamin) mogu da prouzrokuju genotoksične i mutagene efekte. Stoga su ciljevi ovog istraživanja bili da se ispita da li efedrin može da ispolji genotoksične efekte na izolovanim limfocitima čoveka u Komet testu. Odnos doza-efekat određ en je u rasponu koncentracija efedrina od 0.0005 μM do 500 μM. Tri koncentracije efedrina (1, 50 and 300 μM) koje su imale prihvatljiv nivo ćelijske vijabilnosti (preko 90%) upotrebljene su za dalje eksperimente sa inhibitorima reparacije DNK (citozin arabinozid i hidroksiurea). Dobijeni rezultati pokazuju da efedrin nije indukovao oštećenja DNK na izolovanim limfocitima čoveka. Međutim, istovremeni tretman sa inhibitorima reparacije DNK doveo je do malog ali statistički značajnog porasta oštećenja DNK kod negativne kontrole, usled endogenog oštećenja DNK. Interestantno je da ćelije tretirane sa efedrinom i inhibitorima reparacije DNK nisu ispoljile povećan nivo oštećenja DNK. Na osnovu dobijenih rezultata može se zaključiti da efedrin nije ispoljio genotoksične efekte na izolovanim limfocitima čoveka. Ovaj rezultat je u saglasnosti sa prethodnim istraživanjima u kojima je dokazano da efedrin ne dovodi do genotoksičnih efekata u bakterijskim testovima na genske mutacije i u in vitro citogenetičkim analizama.

Premature centromere division (PCD) can be viewed as a manifestation of chromosome instability. In order to evaluate the ability of Paclitaxel (Ptx) and Cycloheximide (Cy) to induce PCD we used a cytokinesis block micronucleus assay (CBMN), fluorescent in situ hybridization (FISH), and the chromosome aberration (CA) assay in human peripheral blood lymphocytes. Results showed that Ptx can induce PCD alone or in combination with Cy. These findings call us to pay more attention to PCD as a parameter of genotoxicity in the pre-clinical research of mono and/or combinational therapies for cancer treatment. Prevremena centromerna deoba (PCD) se može posmatrati kao manifestacija hromozomske nestabilnosti. U cilju procene efekta paklitaksela (Ptx) i cikloheksimida (Cy) na indukciju PCD-a, koristili smo mikronukleus test (CBMN) uz fluorescentnu in situ hibridizaciju (FISH), kao i test hromozomskih aberacija (CA) na humanim limfocitima periferne krvi. Rezultati su pokazali da Ptx sam, ili u kombinaciji sa Cy može da indukuje PCD. Ovi nalazi ukazuju da treba mnogo više pažnje obratiti na pojavu PCD-a kao parametra genotoksičnosti u prekliničkim ispitivanjima mono- i/ili politerapija za lečenje kancera.

Chromosomal alterations are a feature of both aging and Alzheimer's disease (AD). This study examined if premature centromere division (PCD), a chromosomal instability indicator increased in AD, is correlated with aging or, instead, represents a de novo chromosomal alteration due to accelerating aging in AD. PCD in peripheral blood lymphocytes was determined in sporadic AD patients and gender and age-matched unaffected controls. Metaphase nuclei were analyzed for chromosomes showing PCD, X chromosomes with PCD (PCD,X), and acrocentric chromosomes showing PCD. AD patients, regardless of age, demonstrated increased PCD on any chromosome and PCD on acrocentric chromosomes in both genders, whereas an increase in frequency of PCD,X was expressed only in women. This cytogenetic analysis suggests that PCD is a feature of AD, rather than an epiphenomenon of chronological aging, and may be useful as a physiological biomarker that can be used for disease diagnosis.

and physiological effects of adrenaline and other catecholamines, little is known about possible genotoxic and mutagenic effects. There is an interesting experimental finding that dopamine induces DNA strand breaks in human skin fibroblasts and gene mutations in mouse lymphoma cells (M o l d e u s et al., 1983). However, dopamine did not exhibit genotoxic effects in the Salmonella/mammalian-microsome mutagenicity test, the sex-linked recessive lethal test in Drosophila melanogaster, the sister chromatid exchange (SCE) test in human lymphocytes , or the in vivo micronucleus assay in mouse and rat (M o l d e u s et al., 1983). Likewise, in cytogenetic analysis, adrenaline did not exhibit genotoxic effects on cultured human peripheral blood lymphocytes (D j e l i c , 1997). On the other hand, mutagenic effects of various catecholamines on mouse lymphoma cells resulted from the creation of superoxide anion (M c G r e g o r et al., 1988). In addition, catechol derivates, including adrenaline, induce DNA strand breakage by ferryl species, whereas the induction of 8-hydroxyguanine (8OHG) is due to a hydroxyl radical (·OH) (M i u r a et al., 2000). More recent studies revealed that noradrenaline induces primary DNA damage in the Comet assay on purified human lymphocytes (D j e l i c and A n d e r s o n , 2003) and sperm (D o b r z y n s k a et al., 2004), most likely due to formation of reactive oxygen species (ROS). This result is in agreement with findings that adrenaline and other catecholamines can be involved in redox cycling under the influence of superoxide anion (G e n o v a et al., 2006). Interestingly, superoxide anion may induce chromosome breakage and sister chromatid exchange (SCE) (M ’ b e m b a -M e k a et al., 2007).

Alzheimer's disease (AD) is a multi factorial disease, related to the loss of neurons and synapses in cerebral cortex and subcortical structures, leading to degenerative changes and atrophy. Despite abundance of facts related to AD and its pathology, the only drugs used in the prevention and treatment are those from the cholinesterase inhibitors group. However, there is growing evidence that a non-classical therapeutic approach in the treatment of AD has beneficial effects. In this review we summarized recent literature data related to the non-classical drugs for the treatment of AD predominantly used in clinical testing, such as amyloid aggregation inhibitors, beta-sheet breakers, antioxidants, estrogens and immunotherapeutics.

Alzheimer's disease (AD) is a complex and progressive neurodegenerative disorder, and represents the most common form of dementia. The number of people affected by AD is estimated to be doubled by the year of 2050, and more than 100 million people worldwide will be affected by this disease. Still, there is no reliable diagnostic test which would indicate pre-symptomatic conditions or an increased risk of developing AD. The only drugs approved by the FDA belong to the cholinesterase inhibitors (ChEI) group, such as donepezil, rivastigmine, galantamine and memantine that belongs to a class of drugs named receptor NMDA antagonists. Most mainstream pharmacotherapeutic approaches act by slowing the progression of the condition rather than to treat or prevent the cause of AD. In this review we are presenting literature data from recent research related to new avenues in the classical approach to prevention and treatment of AD.