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A. Çavuşoğlu, Zeliha Ipekçi-Altas, K. Bajrovic, N. Gozukirmizi, A. Zehir

Kocaeli University, Arslanbey Agricultural Vocational School, TR-41285 Kartepe, Kocaeli, Turkey. TUBITAK, Research Institute for Genetic Engineering and Biotechnology, TR-41470 Gebze, Kocaeli, Turkey. Institute for Genetic Engineering and Biotechnology, Gajev trg 4/1 BH-71000 Sarajevo, Bosnia and Herzegovnia. Istanbul University, Molecular Biology and Genetics Department, TR-34118 Vezneciler, Istanbul, Turkey. Marmara University, Biology Department, TR-34472 Goztepe, Istanbul, Turkey.

A. Çavuşoğlu, Zeliha Ipekçi-Altas, K. Bajrovic, N. Gozukirmizi, A. Zehir

Populus species are important resource for certain branches of industry and have special roles for scientific study on biological and agricultural systems. Plant regeneration via direct and indirect organogenesis of four Populus deltoides Bartram ex Marsh. ssp. deltoides × Populus deltoides Bartram ex Marsh. ssp. deltoides hybrid clones (89 M 011, 89 M 044, 89 M 048, 89 M 066) and P. deltoides ssp. deltoides clone (Samsun) were investigated. Direct organogenesis was established from nodes and internodes on woody plant medium (WPM) supplemented with cytokinins and/or auxins. The 89 M 011 clone gave the highest percentage (100%) of regeneration on WPM with 1 mg/l zeatine from internode explants. Indirect organogenesis via callus phase was obtained from nodes and petioles on WPM supplemented with different concentrations of 2,4-Dichlorophenoxy acetic acid (2,4-D). The nodes part of the 89 M 066 clone gave the highest rate of generative callus (100%)  on WPM supplemented with 2 mg/l 2,4-D. Indirect shoots were obtained from the node callus on WPM with cytokinins. There was root formation from directly regenerative shoots which were cultured on WPM or Murashige and Skoog Basal Medium (MS) containing different ratios of indole butyric acid (IBA). Rooted seedlings in vitro were successfully acclimatized. Data on in vitro study were subjected to statistical evaluation. The in vitro regeneration system will allow this study to set reliable procedures for the genus and clones. Key words : Poplar, tissue culture, regeneration, organogenesis.

PURPOSE This study assessed the occurrence of Tannerella forsythia in patients with acute and chronic primary endodontic infections. METHODS Clinical samples were collected from 40 patients with acute and chronic periradicular disease. Nested polymerase chain reaction (PCR) assay technique was used to detect the presence of T. forsythia in primary endodontic infections. The first round of PCR amplification used universal primers to detect the 16S rDNA sequence. Product from the first round was then used to amplify T. forsythia specific fragment with species-specific pairs of primers. RESULTS T. forsythia was found in 12 of 27 chronic and 5 of 13 acute infected patients for an overall occurrence frequency of 42.5%. No significant correlation was found between patients with the T. forsythia positive genotype and the occurrence of clinical symptoms in the primary endodontic infections (P < 0.05) (P = 0.496). Also, no significant relationship was found between the occurrence of T. forsythia and the patient's age (P = 0.61) or gender (P = 0.239).

Aim To report on the use of STR, Y-STRs, and miniSTRs typing methods in the identification of victims of revolutionary violence and crimes against humanity committed by the Communist Armed Forces during and after World War II in which bodies were exhumed from mass and individual graves in Slovenia. Methods Bone fragments and teeth were removed from human remains found in several small and closely located hidden mass graves in the Skofja Loka area (Lovrenska Grapa and Žolsce) and 2 individual graves in the Ljubljana area (Podlipoglav), Slovenia. DNA was isolated using the Qiagen DNA extraction procedure optimized for bone and teeth. Some DNA extracts required additional purification, such as N-buthanol treatment. The Quantifiler TM Human DNA Quantifica tion Kit was used for DNA quantification. Initially, PowerPlex 16 kit was used to simultaneously analyze 15 short tandem repeat (STR) loci. The PowerPlex S5 miniSTR kit and AmpFSTR® MiniFiler PCR Amplification Kit was used for additional analysis if preliminary analysis yielded weak partial or no profiles at all. In 2 cases, when the Pow erPlex 16 profiles indicated possible relatedness of the remains with reference samples, but there were insufficient probabilities to call the match to possible male paternal relatives, we resorted to an additional analysis of Y-STR markers. PowerPlex® Y System was used to simultaneously amplify 12 Y-STR loci. Fragment analysis was performed on an ABI PRISM 310 genetic analyzer. Matching probabilities were estimated using the DNA-View software. Results Following the Y-STR analysis, 1 of the “weak matches” previously obtained based on autosomal loci, was confirmed while the other 1 was not. Combined standard STR and miniSTR approach applied to bone samples from 2 individual graves resulted in positive identifications. Finally, using the same approach on 11 bone samples from hidden mass grave Žolosce, we were able to obtain 6 useful DNA profiles. Conclusion The results of this study, in combination with previously obtained results, demonstrate that Y-chromosome testing and mini­ STR methodology can contribute to the identification of human re mains of victims of revolutionary violence from World War II.

AIM To report on the use of STR, Y-STRs, and miniSTRs typing methods in the identification of victims of revolutionary violence and crimes against humanity committed by the Communist Armed Forces during and after World War II in which bodies were exhumed from mass and individual graves in Slovenia. METHODS Bone fragments and teeth were removed from human remains found in several small and closely located hidden mass graves in the Skofja Loka area (Lovrenska Grapa and Zolsce) and 2 individual graves in the Ljubljana area (Podlipoglav), Slovenia. DNA was isolated using the Qiagen DNA extraction procedure optimized for bone and teeth. Some DNA extracts required additional purification, such as N-buthanol treatment. The QuantifilerTM Human DNA Quantification Kit was used for DNA quantification. Initially, PowerPlex 16 kit was used to simultaneously analyze 15 short tandem repeat (STR) loci. The PowerPlex S5 miniSTR kit and AmpF/STR MiniFiler PCR Amplification Kit was used for additional analysis if preliminary analysis yielded weak partial or no profiles at all. In 2 cases, when the PowerPlex 16 profiles indicated possible relatedness of the remains with reference samples, but there were insufficient probabilities to call the match to possible male paternal relatives, we resorted to an additional analysis of Y-STR markers. PowerPlex Y System was used to simultaneously amplify 12 Y-STR loci. Fragment analysis was performed on an ABI PRISM 310 genetic analyzer. Matching probabilities were estimated using the DNA-View software. RESULTS Following the Y-STR analysis, 1 of the "weak matches" previously obtained based on autosomal loci, was confirmed while the other 1 was not. Combined standard STR and miniSTR approach applied to bone samples from 2 individual graves resulted in positive identifications. Finally, using the same approach on 11 bone samples from hidden mass grave Zolosce, we were able to obtain 6 useful DNA profiles. CONCLUSION The results of this study, in combination with previously obtained results, demonstrate that Y-chromosome testing and mini-STR methodology can contribute to the identification of human remains of victims of revolutionary violence from World War II.

Modern Bosnia and Herzegovina is a multi-ethnic and multi-religion country, with a very stormy history. Certain archaeological findings indicate continuous population of its territory since the Paleolithic. In time, vast number of different factors jointly influenced fascinating diversity of local human populations. A great number of small, more or less isolated, indigenous populations, make this area quite attractive for population-genetic surveys of different levels and approaches. Austro-Hungarian military physicians conducted the very first known bio-anthropological analyses of Bosnia-Herzegovina population at the end of the 19th century. Thus, the first step towards resolving the genetic structures of local B&H human populations was made. The studies that followed (conducted throughout most of the 20th century) were primarily based on the observation of various phenotypic traits. This stage was followed by the examination of various cytogenetic and fundamental DNA based molecular markers. The efforts undertaken over the last three centuries revealed "human genetic treasure" in Bosnia and Herzegovina. However, even now, after all the studies that were conducted, many interesting features remain to be discovered and described within the existing local human populations.

AIM To present the joint effort of three institutions in the identification of human remains from the World War II found in two mass graves in the area of Skofja Loka, Slovenia. METHODS The remains of 27 individuals were found in two small and closely located mass graves. The DNA was isolated from bone and teeth samples using either standard phenol/chloroform alcohol extraction or optimized Qiagen DNA extraction procedure. Some recovered samples required the employment of additional DNA purification methods, such as N-buthanol treatment. Quantifiler Human DNA Quantification Kit was used for DNA quantification. PowerPlex 16 kit was used to simultaneously amplify 15 short tandem repeat (STR) loci. Matching probabilities were estimated using the DNA View program. RESULTS Out of all processed samples, 15 remains were fully profiled at all 15 STR loci. The other 12 profiles were partial. The least successful profile included 13 loci. Also, 69 referent samples (buccal swabs) from potential living relatives were collected and profiled. Comparison of victims' profile against referent samples database resulted in 4 strong matches. In addition, 5 other profiles were matched to certain referent samples with lower probability. CONCLUSION Our results show that more than 6 decades after the end of the World War II, DNA analysis may significantly contribute to the identification of the remains from that period. Additional analysis of Y-STRs and mitochondrial DNA (mtDNA) markers will be performed in the second phase of the identification project.

D. Marjanovic, N. Pojskić, J. Davoren, Lejla Kovačević, Adaleta Durmić, N. Bakal, K. Drobnič, D. Primorac et al.

POPULATION: We have analyzed the distribution of allele frequencies at two short tandem repeats loci (D2S1338 and D19S433) in a multinational sample of Bosnia and Herzegovina (B&H) residents. A total of 110 unrelated male and female individuals (Caucasians) from different regions of B&H were sampled for the analysis. We ensured that the sample reflected approximate proportional participation of the three main ethnic groups in the population of B&H (Bosniacs‐Muslim [45%], Serbs [34%], Croats [21%]).

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