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P. Priftakis, T. Dalianis, J. Carstensen, U. Samuelsson, I. Lewensohn‐Fuchs, G. Bogdanovic, J. Winiarski, B. Gustafsson

BACKGROUND Epidemiological evidence has suggested that some childhood acute lymphoblastic leukemia (ALL) may be initiated in utero and may have an infectious etiology. The human polyomavirus JC virus (JCV) has been discussed as a candidate virus, but its presence has not been demonstrated in leukemia cells from children with ALL. The aim of this study was, therefore, to investigate if prenatal human polyomavirus infection could still indirectly be correlated to the development of childhood ALL. PROCEDURE Fifty-four Guthrie cards (stored, dried blood spots filter papers, routinely collected from newborns for different screening analyses), collected at 3-5 days of age, from Swedish children who subsequently developed ALL, as well as from 37 healthy controls, were investigated by nested PCR for the presence of human polyomaviruses JCV and BK virus (BKV). RESULTS JCV and BKV DNA were not detected in any of the Guthrie cards from ALL patients or from healthy controls, although all tested samples had amplifiable DNA as confirmed by an HLA DQ PCR. CONCLUSIONS JCV or BKV were not found in any of the dried blood spots of children who later developed ALL or in the healthy controls. These findings suggest that it is unlikely that childhood ALL is associated with an in utero infection with JCV or BKV, although it is not possible to exclude an association with an in utero infection that has become latent in the kidneys with very low levels of circulating virus at birth.

INTRODUCTION Preterm labour is most common complication of second half of pregnancy, incidence is 7 to 10% of all delivers. One of most important causes of increased mortality and morbidity of neonates is respiratory distress syndrome. Numerous studies prove that corticosteroids given antenatal to mother decrease the incidence of preterm delivered neonates. AIM OF THIS STUDY IS: To determined influence of dexamethasone given prepartal on maturation of neonatal lungs in correlation with gestational age. METHOD This study include 150 pregnant women which delivered before 37 week of gestation. They are divide in three groups: two experimental and one control group. Group E1 consists of the pregnant women which received dexamethasone for five days in a single dose of 12 mg. Group E2 consists of the pregnant women which received dexamethasone less then five days, in a single dose of 12 mg. In the control group consists of the pregnant women which did not received dexamethasone. In this work we used gestational age in the moment of delivery. State of neonate is determined on the base of presence of clinical signs of respiratory distress syndrome after birth. Statistical method used in this work was test of proportion. RESULTS In this work we find that there is a less number of neonates who has respiratory distress syndrome in group consists of the pregnant women which received dexamethasone for five days, 5 (10%). In group consists of the pregnant women which received dexamethasone less then five days, number of neonates with respiratory distress syndrome was 12 (24%). The number of neonates with respiratory distress syndrome in control group was 21 (42%). The value of test of proportion: K/E1 (the pregnant women received dexamethasone for five days)-z = 1.95, p < 0.05; K/E2 (pregnant women received dexamethasone less than five days)-z = 3.92, p < 0.05. In all three groups largest number of neonates with respiratory distress syndrome was between 31-34 week of gestation. Highest mortality of neonate with respiratory distress syndrome was in control group, 7 (14%), than in group consists of the pregnant women received dexamethasone lass then five days, 4 (8%). In group consist pregnant women received dexamethasone for five days, there were not cases of mortality caused by respiratory distress syndrome. The value of test of proportion was z = 2.85 (p < 0.05), between control group and group consists of the pregnant women received dexamethasone for five days. CONCLUSION Dexamethasone accelerates maturation of fetal lungs, decrease number of neonates with respiratory distress syndrome and improves survival in preterm delivered neonates. Optimal gestational age for use of dexamethasone therapy is 31 to 34 weeks of gestation.

P. Priftakis, G. Bogdanovic, A. Hjerpe, T. Dalianis

UNLABELLED Simian virus 40 (SV40), a contaminant of polio vaccines used in the United States and Europe between 1955 and 1963, has been detected with high frequency in human malignant mesotheliomas (MM). In Sweden, from 1958, due to production in Javanese macaque kidney cells and SV40 testing from 1961, only polio vaccine claimed to be free of SV40 has been used. Hence, we aimed to screen Swedish MM patients for the presence of SV40. MATERIALS AND METHODS Forty-one paraffin-embedded pleural MM samples, obtained from patients born from 1893-1958, were examined for amplifiable DNA. Testable samples were thereafter evaluated for the presence of SV40 DNA by two types of polymerase chain reactions (PCR) followed by sequencing. RESULTS SV40 could be confirmed by sequencing in only 3 of the 30 MM samples containing DNA that could be amplified by PCR. CONCLUSION The presence of SV40 is not frequent in Swedish MM patients.

G. Bogdanovic, V. Kojić, T. Srdić, D. Jakimov, M. Djuran, Ž. Bugarčić, M. Baltić, V. Baltic

The platinum (II)complexes, cis-[PtCl2(CH3SCH2CH2SCH3)] (Pt1), cis-[PtCl2(dmso)2] (dmso is dimethylsulfoxide; Pt2) and cis-[PtCl2(NH3)2] (cisplatin), and taxol (T) have been tested at different equimolar concentrations. Cells were exposed to complexes for 2 h and left to recover in fresh medium for 24, 48 or 72 h. Growth inhibition was measured by tetrazolium WST1 assay Analyses of the cell cycle, and apoptosis were performed by flow cytometry, at the same exposure times. The IC50 value of each platinum(II) complex as well as combination index (CI; platinum(II) complex + taxol) for various cytotoxicity levels were determined by median effects analysis. MCF7 cells were found to be sensitive to both Pt1 and Pt2 complexe These cisplatin analogues influenced the cell growth more effectively as compared to cisplatin. Cytotoxic effect was concentration and time-dependent. Profound growth inhibitory effect was observed for Pt1 complex, across all its concentrations at all recovery periods. A plateau effect was achieved three days after treatment at Pt1 concentrations ≤ 1 μM. Pt2, however, decreased MCF7 cells survival only for the first 24 h ranging between 50-55%. Pt2 cytotoxicity sharply decreased thereafter, approaching 2 h - treatment cytotoxicity level. The median IC50 values for Pt1 and Pt2 were similar (0.337 and 0.3051 μM, respectively) but only for the first 24 h. The IC50 values for Pt1 strongly depend on the recovery period. On simultaneos exposure of cells to taxol and platinum(II) complexes no consistent effect was found. The Cls for combinations of taxol with Pt1 or Pt2 revealed cytotoxic effects that were in most Cases synergistic (Pt1) or less than addtiive (Pt2). Flow cytometry analysis has shown that each platinum(II) complex induced apoptosis in MCF7 cells. The level of apoptosis correlated with cytotoxicity level for the range concentrations. Both cisplatin analogues, at IC50 concentrations, increased the number of MCF7 cells in G0G1 phase of cell cycle. Pt2-treated cells remained arrested in G0G1 phase up to 72 h after treatment. Combination of Pt2 and taxol caused further arrest of cells in G0G1 phase (24 h) in parallel with strong decrement of G2M phase cells.

V. Jurišić, G. Bogdanovic, T. Srdić, N. Kraguljac, M. Baltić, V. Baltic

V. Jurišić, G. Bogdanovic, T. Srdić, N. Kraguljac, A. Kerenji, M. Baltić, V. Baltic

V. Jurišić, G. Bogdanovic, T. Srdić, N. Kraguljac, A. Kerenji, M. Baltić, V. Baltic

A. Đorđević, M. Vojinovic-Miloradov, G. Bogdanovic

This paper presents the basic chemical characteristics of buckminsterfullerene-molecule CM60 and some of its biologically active derivatives. This new form of carbon with unique properties, discovered in 1985, is a very reactive species undergoing a variety of chemical reactions leading to a wide spectrum of possible covalent derivatives. Besides covalent compounds, fullerene reacts with different dopants to form non-covalent complexes, including endohedral, exohedral systems (intercalates). The review of the most significant papers on noncovalent and covalent derivatives and their biological activity is given, as well as our results in this field. Formation of intercalates of fullerene with small molecules of aliphatic alcohols is described and the importance of topology and size of dopants and the role of solvents in formation of intercalates is explained. Original synthesis of bromine derivative C60Br24 and water-soluble polyhydroxy derivative C60(OH)24 is described. Antioxidative effects of the latter compound were investigated in chemical model system and in vitro system of human neoplasmatic cells. The paper also gives theoretical explanation of addition C60R24 derivatives together with assumption about antioxidative activity of fullerol. .

J. Zdravković, M. Avramov-Ivić, D. Vuković, G. Bogdanovic, U. Mioč, D. Četojević-Simin

P. Priftakis, G. Bogdanovic, G. Tydén, T. Dalianis

ABSTRACT Polyomaviruria was observed in one-third of all renal transplant patients, irrespective of whether their renal grafts came from a living or cadaver donor, and was not correlated to graft rejection episodes. This suggests that the renal graft ischemia period is not the major cause of polyomavirus reactivation and that reactivation of polyomavirus is not a dominant cause of graft rejection.

G. Bogdanovic, P. Priftakis, B. Taemmeraes, A. Gustafsson, T. Flægstad, J. Winiarski, T. Dalianis

In allogeneic bone marrow transplanted (BMT) patients BK virus (BKV) reactivation has been associated with haemorrhagic cystitis (HC). However, it is far from obvious which patients will develop HC, since BKV, a human polyomavirus, is ubiquitious and infects children at an early age. To investigate if a primary BKV infection, as such or possibly due to transmission of BKV by the marrow graft during BMT, was correlated to the development of HC, 45 children were followed for possible BKV seroconversion and development of HC at different time points after BMT. Serum samples were collected from the 45 allogeneic BMT children and their donors before transplantation, and from the patients at 3, 6 and 12 months after BMT. These sera were analysed for the presence of specific antibodies towards BKV by hemagglutination inhibition (HAI) and by IgG- and IgM-class specific enzyme linked immunosorbent (ELISA) assays. Twelve of the 45 BMT children had a documented episode of HC or hematuria. All patients and 98% of the donors were HAI positive before BMT, while with ELISA 87% of the patients and 84% of the donors were positive. Moreover, most HC and hematuria children (11/12) were seropositive with both assays before BMT, making it impossible to investigate possible BKV transmission through the bone marrow graft during BMT by serology. Still, serological changes such as ELISA seroconversion, IgM antibodies and/or HAI titer increases were significantly (p=0.016) more common in patients with HC (58%) than without HC (24%), but these changes occured mainly after HC symptomatology had already resolved. However, there was a near significant difference (p=0.053) in BKV seroprevalence by ELISA among the donors of patients with HC or hematuria (67%) as compared to the donors (91%) of patients without HC.

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