Sporadic chromosome translocation frequencies in lymphocyte cultures – a retrospective study in a cohort of patients from Bosnia and Herzegovina

Aim Chromosome translocations are considered as one of the most severe forms of genome defects. Because of the clinical significance of chromosome translocations and scarce data on the incidence of sporadic translocations in population of Bosnia and Herzegovina, we aimed to report sporadic translocation frequencies in samples karyotyped in our laboratory. Methods The study group consisted of 108 samples. Whole blood was cultivated in complete medium for 72 hours with the thymidine application at 48th hour to synchronize the cell culture. Metaphases were arrested by colcemid 60 minutes before harvesting. Following hypotonic treatment, cells were fixed and cell suspension was dropped on coded slides. Dried slides were subjected to conventional GTG (G-banding with trypsin-Giemsa) banding and analyzed under 1000x magnification in the accordance with ISCN (International System for Human Cytogenetic Nomenclature) and E.C.A. Cytogenetic Guidelines and Quality Assurance. Results The incidence of all detected sporadic translocations was 27.81 x 10-4 per metaphase. The incidence of sporadic translocations involving chromosomes 7 and 14, being considered as the most frequent sporadic translocations of the human karyotype in phytohaemagglutinin (PHA) stimulated lymphocytes, was 15.89 x 10-4 per metaphase. The most frequent breakpoints were 7p21, 14q11 and 14q21. Other detected sporadic translocation breakpoints were: 1q25, 3p22, 7p13, 7q11.22, 7q33, 14q23 and 19q13.4. Conclusion Higher incidence of sporadic translocations compared to the similar studies was registered. Since potential explanations for this issue are smaller sample size and higher exposure of examined population to genotoxic agents, further monitoring of sporadic translocation incidences is recommended

A retrospective analysis of spontaneous chromosomal aberrations in human lymphocyte cultures of individuals from Bosnia and Herzegovina

Spontaneous chromosomal aberrations are structural or numerical changes of chromosomes that occur naturally, without exposure to external genotoxic factors. They are not inherited, occur randomly in the karyotype, and do not have direct clinical significance. However, they can affect genomic instability and disease predisposition. They can result from DNA replication or repair processes errors, and typically are observed in cells that are actively dividing. Spontaneous chromosomal aberrations may arise due to the natural chromosomal instability and can be elevated in individuals exposed to mutagens. We analyzed frequencies of spontaneous chromosomal aberrations in 137 individuals subjected to karyotype analysis at the Laboratory for Cytogenetics and Genotoxicology, University of Sarajevo – Institute for Genetic Engineering and Biotechnology, during 2008-2023. Whole blood samples were cultivated for 72 hours with the thymidine added in the 48th hour. Metaphases were arrested by colcemid 60 minutes before harvesting. GTG banding was performed and slides were analyzed under 1000x magnification in accordance with An International System for Human Cytogenetic Nomenclature and E.C.A. Cytogenetic Guidelines and Quality Assurance. Constitutionally aberrant karyotypes were found in 2.92% of analysed individuals as well as altered karyotypes considered as normal chromosomal variants. In the total of 3092 analyzed metaphases, 20 spontaneous chromosomal aberrations were found in 13 individuals. This study contributes to the limited knowledge of the cytogenetic status of the Bosnian and Herzegovinian population. Further monitoring of spontaneous chromosomal aberrations incidences is recommended

Razvoj novih antidijabetičkih lekova na bazi polioksometalatnih nanoklastera

Zahvaljujući brojnim istraživanjima koja su ispitivala biološka svojstva strukturno različitih polioksometalata, došlo se do zapažanja da ova kompleksna neorganska jedinjenja, pored antimikrobnog i antitumorskog delovanja, mogu biti delotvorna u snižavanju hiperglikemije kod pacova sa eksperimentalno izazvanim dijabetesom. Stoga, cilj ove studije je bio da se ispitaju antidijabetički potencijal i mogući toksični efekti dva polioksovolframata:(NH4 )14[NaP5 W30O110]·31H2 O, {NaP5 W30} i K14[AgP5 W30O110]·22H2 O·6KCl, {AgP5 W30}. U cilju realizacije postavljenog cilja, korišćena su tri eksperimentalna modela: (1) antihiperglikemijska screening studija u kojoj je ispitivan uticaj jednokratne intraperitonealne primene {NaP5 W30} i {AgP5 W30} (5, 10 i 20 mg/kg) na snižavanje hiperglikemije kod dijabetičkih pacova, (2) akutna peroralna toksikološka studija koja je istraživala hepato- i nefrotoksične efekte odabranih heteropolivolframata kod zdravih pacova i (3) studija posvećena rasvetljavanju mogućih mehanizama antidijabetičkog delovanja heteropolivolframata. Rezultati screening studije su pokazali da su oba ispitivana heteropolivolframata efikasna u snižavanju hiperglikemije, s tim što se {NaP5 W30}, u odnosu na {AgP5 W30}, pokazao kao moćniji antihiperglikemijski agens. Rezultati biohemijskih parametara funkcije i patohistološka analiza jetre i bubrege korišćenjem konvencionalne svetlosne i transmisione elektronske mikroskopije pokazuju da dvonedeljna primena {NaP5 W30} i {AgP5 W30} (20 mg/kg) izaziva blagi do umereni stepen hepato- i nefrotoksičnosti kod zdravih životinja. U poslednjem eksperimentalnom protokolu, pokazano je da tronedeljna peroralna primena {NaP5 W30} (20 mg/kg) povećava koncentraciju insulina u serumu dijabetičkih pacova, što može biti jedan od mehanizama njegovog antidijabetičkog delovanja. Takođe, pokazano je da {NaP5 W30} ispoljava hepato-, nefro-, kardio- i neuroprotektivno dejstvo kod dijabetičkih pacova, što je procenjeno na osnovu analize: (1) relativne mase organa, (2) biohemijskih parametara funkcije, (3) parametara oksidativnog stresa u homogenatu tkiva, (4) aktivnosti acetilholinesteraze, Na+ /K+-ATPaze i ecto-ATPaza u sinaptozomima i (5) patohistoloških promena u tkivima korišćenjem konvencionalne svetlosne i transmisione elektronske mikroskopije. Stoga, {NaP5 W30} i {AgP5 W30} mogu se smatrati mogućim neinsulinskim lekovima-kandidatima u terapiji dijabetesa tipa 2, koji bi se podvrgli daljim pretkliničkim istraživanjima.Simpozijum „Stremljenja i novine u medicini“ Medicinskog fakulteta u Beogradu, Beograd, 04-08. decembra, 2023

Monolacunary Wells-Dawson Polyoxometalate as a Novel Contrast Agent for Computed Tomography: A Comprehensive Study on In Vivo Toxicity and Biodistribution

Polyoxotungstate nanoclusters have recently emerged as promising contrast agents for computed tomography (CT). In order to evaluate their clinical potential, in this study, we evaluated the in vitro CT imaging properties, potential toxic effects in vivo, and tissue distribution of monolacunary Wells–Dawson polyoxometalate, α2-K10P2W17O61.20H2O (mono-WD POM). Mono-WD POM showed superior X-ray attenuation compared to other tungsten-containing nanoclusters (its parent WD-POM and Keggin POM) and the standard iodine-based contrast agent (iohexol). The calculated X-ray attenuation linear slope for mono-WD POM was significantly higher compared to parent WD-POM, Keggin POM, and iohexol (5.97 ± 0.14 vs. 4.84 ± 0.05, 4.55 ± 0.16, and 4.30 ± 0.09, respectively). Acute oral (maximum-administered dose (MAD) = 960 mg/kg) and intravenous administration (1/10, 1/5, and 1/3 MAD) of mono-WD POM did not induce unexpected changes in rats’ general habits or mortality. Results of blood gas analysis, CO-oximetry status, and the levels of electrolytes, glucose, lactate, creatinine, and BUN demonstrated a dose-dependent tendency 14 days after intravenous administration of mono-WD POM. The most significant differences compared to the control were observed for 1/3 MAD, being approximately seventy times higher than the typically used dose (0.015 mmol W/kg) of tungsten-based contrast agents. The highest tungsten deposition was found in the kidney (1/3 MAD—0.67 ± 0.12; 1/5 MAD—0.59 ± 0.07; 1/10 MAD—0.54 ± 0.05), which corresponded to detected morphological irregularities, electrolyte imbalance, and increased BUN levels

Monolacunary Wells-Dawson Polyoxometalate as a Novel Contrast Agent for Computed Tomography: A Comprehensive Study on In Vivo Toxicity and Biodistribution

Polyoxotungstate nanoclusters have recently emerged as promising contrast agents for computed tomography (CT). In order to evaluate their clinical potential, in this study, we evaluated the in vitro CT imaging properties, potential toxic effects in vivo, and tissue distribution of monolacunary Wells–Dawson polyoxometalate, α2-K10P2W17O61.20H2O (mono-WD POM). Mono-WD POM showed superior X-ray attenuation compared to other tungsten-containing nanoclusters (its parent WD-POM and Keggin POM) and the standard iodine-based contrast agent (iohexol). The calculated X-ray attenuation linear slope for mono-WD POM was significantly higher compared to parent WD-POM, Keggin POM, and iohexol (5.97 ± 0.14 vs. 4.84 ± 0.05, 4.55 ± 0.16, and 4.30 ± 0.09, respectively). Acute oral (maximum-administered dose (MAD) = 960 mg/kg) and intravenous administration (1/10, 1/5, and 1/3 MAD) of mono-WD POM did not induce unexpected changes in rats’ general habits or mortality. Results of blood gas analysis, CO-oximetry status, and the levels of electrolytes, glucose, lactate, creatinine, and BUN demonstrated a dose-dependent tendency 14 days after intravenous administration of mono-WD POM. The most significant differences compared to the control were observed for 1/3 MAD, being approximately seventy times higher than the typically used dose (0.015 mmol W/kg) of tungsten-based contrast agents. The highest tungsten deposition was found in the kidney (1/3 MAD—0.67 ± 0.12; 1/5 MAD—0.59 ± 0.07; 1/10 MAD—0.54 ± 0.05), which corresponded to detected morphological irregularities, electrolyte imbalance, and increased BUN levels

In vivo toxicity evaluation of a polyoxotungstate nanocluster as a promising contrast agent for computed tomography

Abstract In this study, we demonstrate for the first time, that a discrete metal-oxo cluster α-/β-K6P2W18O62 (WD-POM) exhibits superior performance as a computed tomography (CT) contrast agent, in comparison to the standard contrast agent iohexol. A toxicity evaluation of WD-POM was performed according to standard toxicological protocols using Wistar albino rats. The maximum tolerable dose (MTD) of 2000 mg/kg was initially determined after oral WD-POM application. The acute intravenous toxicity of single WD-POM doses (1/3, 1/5, and 1/10 MTD), which are at least fifty times higher than the typically used dose (0.015 mmol W kg−1) of tungsten-based contrast agents, was evaluated for 14 days. The results of arterial blood gas analysis, CO-oximetry status, electrolyte and lactate levels for 1/10 MTD group (80% survival rate) indicated the mixed respiratory and metabolic acidosis. The highest deposition of WD-POM (0.6 ppm tungsten) was found in the kidney, followed by liver (0.15 ppm tungsten), for which the histological analysis revealed morphological irregularities, although the renal function parameters (creatinine and BUN levels) were within the physiological range. This study is the first and important step in evaluating side effects of polyoxometalate nanoclusters, which in recent years have shown a large potential as therapeutics and contrast agents

In vivo toxicity evaluation of two polyoxotungstates with potential antidiabetic activity using Wistar rats as a model system

In this study, thein vivohypoglycemic effect of a donut-shaped polyanion salt (NH4)14[Na@P5W30O110]$31H2O{NaP5W30} and its Ag-containing derivative K14[Ag@P5W30O110]$22H2O$6KCl {AgP5W30}, as wellas their hepatotoxicity and nephrotoxicity, was evaluated. In the screening hypoglycemic study,Wistaralbinorats with streptozotocin induced diabetes were treated intraperitoneally with three single doses (5,10, and 20 mg per kg per b.w.) of both investigated polyoxotungstates. The blood glucose levels,measured before and after 2, 4 and 6 h polyoxotungstate application, showed that both studiedcompounds induced the most pronounced and time dependent glucose lowering effects at the doses of20 mg kg1. Thus, daily doses of 20 mg kg1were administered toWistar albinorats orally for 14 days infurther toxicity examinations. The serum glucose concentration and biochemical parameters of kidneyand liver function, as well as a histopathological analysis of kidney and liver tissues were evaluated 14days after the polyoxotungstate administration. Both investigated compounds did not induce statisticallysignificant alterations of the serum glucose and uric acid concentrations, as well as some of the liverfunction markers (serum alanine and aspartate aminotransferases, and alkaline phosphatase activities).However, the significant decrease in serum total protein and albumin concentrations and the increase inbiochemical parameters of renal function–serum urea (up to 63.1%) and creatinine concentrations (upto 23.3%) were observed for both polyoxotungstates. In addition, the detected biochemical changeswere in accordance with kidney and liver histhopathological analysis. Accordingly, the hepatotoxic andnephrotoxic effects of these potential antidiabetic polyoxotungstates could be considered as mild