Maciej Wnuk, Monika Bugno-Poniewierska, Anna Lewińska, Bernadetta Oklejewicz, Tomasz Ząbek and Ewa Słota
Aging Process in Chromatin of Animals
The aging process is a variable, stochastic and pleiotropic phenomenon which is regulated by different environmental and genetic factors. The age-associated changes, which occur at the molecular and cellular levels and disturb biological homeostasis, may directly or indirectly contribute to aging, causing apoptosis or cellular senescence and consequently leading to the death of the organism. In this context, it is particularly interesting to observe changes in somatic cell chromatin. In the present paper, we summarized the knowledge on the biological aspects of aging with special consideration of age-related changes in chromatin like DNA damage, shortening telomeres or age-related changes in methylation of DNA.
Małgorzata Nadziakiewicz, Halina Kurzawińska, Stanisław Mazur and Dorota Tekielska
Symptoms of discoloration and necrosis of the leaves/needles and shoots of plants are an increasingly common phenomenon in nurseries. They necessitate the withdrawal of the affected plants from sale, which has significant economic consequences. In 2010-2011, observations were conducted of the health of shrubs in nurseries of the Małopolska province. Disease symptoms were mostly found in juniper (Juniperus horizontalis ‘Wiltonii’), rose (ground-cover rose ‘Star Profusion’), yew (Taxus × media ‘Hillii’) and highbush blueberry (Vaccinium corymbosum ‘Patriot’). These species were selected for further study. The affected shrubs represented more than 46% of the population of a given species. Fragments of the diseased organs: the leaves or needles, the base of the shoots, and the roots, were collected from the borderline between healthy and diseased tissue, and used to isolate and identify the microorganisms colonizing the diseased parts. The affected organs were found to be inhabited to the largest extent by the fungus Alternaria alternata (Fr.) Keissl., which accounted for 19.7 to 47.5% of the isolates from the tested species of shrubs. There were also large populations of fungi of the genera: Botrytis (up to 9.82%), Cladosporium (up to 5.66%), Colletotrichum (up to 5.13%), Fusarium (up to 18.38%), Mortierella (up to 7.26%), Pestalotia (up to 5.36%), Rhizoctonia (up to 5.36%), Sclerotinia (up to 6.99%), and Trichoderma (up to 17.09%). The fungus A. alternata, being by far the dominant pathogen, was tested for its pathogenicity for the shoots of the chosen species of shrubs. The test was conducted for 14 days in a chamber with parameters so programmed that they reflected the natural conditions at the height of the growing season. The fungus A. alternata exhibited pathogenicity for all of the tested species of shrubs. Necrosis developed on all the inoculated fragments of shoots. The surface area of necrotic lesions was larger on the shoots of juniper and blueberry.
The evolution of modern humans is a long and difficult process which started from their first appearance and continues to the present day. The study of the genetic origin of populations can help to determine population kinship and to better understand the gradual changes of the gene pool in space and time. Mitochondrial DNA (mtDNA) is a proper tool for the determination of the origin of populations due to its high evolutionary importance. Ancient mitochondrial DNA retrieved from museum specimens, archaeological finds and fossil remains can provide direct evidence for population origins and migration processes. Despite the problems with contaminations and authenticity of ancient mitochondrial DNA, there is a developed set of criteria and platforms for obtaining authentic ancient DNA. During the last two decades, the application of different methods and techniques for analysis of ancient mitochondrial DNA gave promising results. Still, the literature is relatively poor with information for the origin of human populations. Using comprehensive phylogeographic and population analyses we can observe the development and formation of the contemporary populations. The aim of this study was to shed light on human migratory processes and the formation of populations based on available ancient mtDNA data.
Tomasz Ząbek, Ewelina Semik, Agnieszka Fornal, Artur Gurgul and Monika Bugno-Poniewierska
One of epigenetic features of mammalian genomes is methylation of DNA. This nucleotide modification might exert suppressive effect on gene transcription. We have described putative relevance of methylation of one of immune cells related gene (ITGAL) observed in the set of 11 equine tissues. Comparison between qualitative RT-PCR results and DNA bisulfite sequencing of investigated set of tissues pointed to potential correlations between tissue specific methylation and tissue specific transcription in ITGAL locus. These findings might be important for studies on genetic and epigenetic background of autoimmune disorders in the horse.
The guanaco (Lama guanicoe) is the major inhabitant and the largest wild artiodactyl in Patagonia. The introduction of invasive species into its ecological niche poses ecological risks, since invasive species may introduce harmful parasites to this native species. In this work, filariform larvae of the Nematodirus genus were found in feces of guanacos from the Perito Moreno National Park in Argentina. All species were characterized according to morphological features and molecular analyses using ribosomal DNA (rDNA). For the molecular analysis, rDNA fragments were amplified by PCR and then sequenced. The results of the BLASTN comparison threw a 99 % of identity with Nematodirus spathiger and 97 % with N. helvetianus, suggesting that N. spathiger is the infecting parasite. Nematodirus spathiger together with N. filicollis and N. battus causes diarrhea and deaths in sheep and, in some cases, in South American camelids. The availability of more accurate diagnostic methods such as PCR could improve the control measures for gastrointestinal helminthiasis.
Saša Marić, Andrej Razpet, Vera Nikolić, Aleš Snoj and Predrag Simonović
The purpose of this study was to assess the genetic diversity of Serbian huchen and provide guidelines for conservation and management. Fourteen huchen individuals were collected from six localities. For all individuals the mitochondrial DNA control region and the NADH1 gene were sequenced, and two microsatellite loci were genotyped. Sequencing of two mtDNA loci showed the presence of the same haplotypes (Hh_CR_1 and Hh_ND1_1) in all investigated specimens and the complete absence of genetic variability at inter- and intra-populational level. However, analyzed individuals from Serbia have the largest number of control region repeats (average 8.7) compared to all other huchen populations from the Danube basin. Microsatellite data show that the most of Serbian huchens were placed in the “eastern” cluster except one individual from the Drina River which clustered with “western” samples, most likely a consequence of stocking. In spite of only two loci used, four unique alleles found in the studied Serbian and Drina basin population indicate that the Danube basin may possess more region specific alleles than previously thought, which could be of great importance for efficient species management in the future.
Longidorus caespiticola was found for the first time from the rhizosphere of apple with two other Longidorus sp. at Žlunice, Czech Republic. Females and males were analyzed morphologically and morphometrically. Four regions (18S, ITS1, ITS2, D2-D3 expansion segments of 28S) of ribosomal DNA and one region (cox1) of mitochondrial DNA were sequenced and analyzed and compared with a population from the Slovak Republic. The partial mitochondrial cytochrome c oxidase subunit 1 (cox1) gene showed relatively high genetic variation compared with ribosomal DNA between the Czech and Slovakian populations.
Loredana Mariana Aftenie, Irina Franciuc, Alina Martinescu and Adina Honcea
Obtaining blood biospecimens presents logistical and financial challenges. As a result, saliva biospecimen collection is becoming more frequent because of the ease of collection and lower cost. This article describes an assessment of two different methods for collecting samples: whole blood and whole saliva samples used further for DNA extraction and HLA genotyping in immunogenic disease on a group of patients registered at our Molecular Genetics Laboratory Faculty of Medicine “Ovidius” University Constanţa. Our data show that only 81% of the requested participants delivered a blood sample, whereas 19% delivered a saliva sample because they refuse the first sampling method. Analysis of purified genomic DNA by Nano Photometer and agarose gel electrophoresis revealed that blood and saliva samples resulted in DNA with the best quality. PCR analysis showed that DNA from 100% of the blood samples and 93% of the saliva samples could be subsequently amplified. Our study shows that the response rate of self-collection saliva samples had to be considering for the patients that have a low response rate of blood sampling. The quality of genomic DNA from saliva samples was comparable with blood samples as assessed by purity, concentration, yield and PCR amplification. We conclude that the use of saliva samples is a good alternative to blood samples to obtain genomic DNA of high quality and it will considerably increase the participant’s response rate for genetic studies.
Mitochondrial DNA 4977 bp Deletion in Chronic Cervicitis and Cervix Cancers
Mitochondrial DNA (mtDNA) mutations have been implied in many diseases including cancer and inflammatory diseases. The aim of this study is to investigate the relationship between the 4977 bp deletion of the mtDNA and chronic cervicitis or cervix cancer in patients. The study included a group of patients with chronic cervicitis or cervix cancer, and a control group consisting of individuals without any cervical tissue disease. A total of 72 subjects in an East Turkish population were included in the study. Of these, 35 had chronic cervicitis, 21 had cervix cancer and 16 served as the control group. Isolation of mtDNA was performed from the tissues of these patients and then mtDNA deletions were studied using polymerase chain reaction (PCR). In the cancer groups, there were 9.5% heteroplasmic and homoplasmic deletions. There were no homoplasmic deletions in the cervicitis and control groups, but the frequencies of heteroplasmic deletions were 80.0 and 31.2%, respectively. Chronic inflammation leading to increased reactive oxygen species (ROS) may be the cause of the high mtDNA 4977 bp deletion frequencies in cancer and cervicitis. The older age of the cancer patient may suggest that ageing in addition to long time exposure to ROS may lead to deletions and subsequently cancer. This is the first study to investigate the relationship of the mtDNA 4977 bp deletion to chronic cervicitis and cervix cancer.
Alexandros Papachristoforou, Agnès Rortais, Maria Bouga, Gérard Arnold and Lionel Garnery
Honey bee populations from the island of Cyprus were analyzed using microsatellite and mitochondrial DNA markers. A total of 268 colonies were sampled in Cyprus, at six different locations-Kyrenia, Katydata, Flassou, Alabra, Troulloi, and Alassa-covering a wide area of the island. Results showed that the Cyprian honey bee Apis mellifera cypria could be distinguished from other Apis mellifera subspecies based on a „double pattern“ of mitochondrial DNA belonging to the C1 lineage and microsatellite DNA belonging to the O lineage. All populations were homogeneous, except the population from Kyrenia, probably due to the introduction of queens or colonies belonging to the C2, C6, and M7 lineages.