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This article continues a historical outline of the rural municipality administration and local self-government in rural China and Taiwan. The authors analyze volost administration and local self-government in a modern Chinese village.

Taking into account the latest changes in legislation, the monograph examines the norms of administrative-tort and criminal legislation providing for administrative and criminal liability for illegal archaeological activities in the Russian Federation. According to the author, the emerging judicial practice in Russia in cases of administrative offenses provided for in Articles 7.15, 7.15.1 of the Administrative Code of the Russian Federation, and criminal cases of a crime provided for in Article 243.2 of the Criminal Code of the Russian Federation raises many questions, which requires further improvement of Russian legislation in the field of protection of cultural heritage objects. The author suggests a number of innovations that could be taken into account when developing a law on archaeological heritage sites and on improving the relevant protective norms. Special attention is paid to the analysis of norms-definitions that reveal the content of the basic concepts: "archaeological heritage object", "archaeological objects", "cultural layer". The necessity of abandoning the "floating" age criterion (at least one hundred years old), fixed in the Federal Law "On Objects of Cultural Heritage (historical and cultural monuments) of the peoples of the Russian Federation", is substantiated. Instead of the age criterion, it is proposed to introduce the concept of "archaeological epoch" into legal circulation. The content of the concepts is also revealed: "illegal search and seizure of archaeological objects from the archaeological cultural layer", "archaeological cultural layer", "damage to the archaeological cultural layer", "destruction of the archaeological cultural layer". It is intended for scientists and practitioners, law enforcement officials, public authorities and local self-government, teachers, graduate students and students of law schools and faculties, historians, as well as anyone interested in the legal protection of archaeological heritage sites.

The main task in the development of concentrating systems is in reducing the inhomogeneous distribution of illumination over the focal spot, which determines the distribution of concentrated radiation on the receiver, which affects the efficiency of heat and photo converters. (Research purpose) The research purpose is in modeling for calculation, construction manufacturing and research of characteristics of a thermal photovoltaic module with specified energy parameters. (Materials and methods) The article considers new thermal photovoltaic modules that include photodetectors and concentrators that provide efficient conversion of solar energy into heat. The research is based on mathematical modeling techniques using the laws of photoelectricity, geometric optics, and heat and mass transfer. The computer-aided design system COMPASS-3D was used to create a prototype model of the solar module, and the FDM 3D printing technology was used to produce the resulting CAD models. Authors conducted bench and field tests of components and complexes of solar power systems in order to clarify the design parameters. (Results and discussion) During the study of a developed solar module with parabolic concentrators and a discrete photodetector with asymmetric profile, with a system of the coolant flow, physical and mathematical models on the basis of which was calculated: the structural parameters; the distribution of concentration of solar radiation across the width of the photodetector; dependences of the distribution of the heating temperature of the coolant over the profile of the photodetector. Authors designed a layout of the basic thermal photovoltaic module, in which the rounded lower part of the rotary rack fixed on the axis allows to create a rotation in the axial direction of the photodetector holders within ±20 degrees with a mechanism for fixing the adjusted position. (Conclusions) Based on the alignment carried out on the stand, the identical location of the receivers of concentrated solar radiation relative to the concentrators in the installation is preserved.

We have previously reported that depletion Cyclophilin D (CypD), a regulatory subunit of mitochondrial permeability transition pore, improves vascular function and attenuates hypertension, however, specific regulation of CypD in hypertension is not clear. Analysis of human arterioles from hypertensive patients did not reveal alterations in CypD levels but showed 3-fold increase in CypD acetylation. We hypothesized that CypD-K166 acetylation promotes vascular oxidative stress and hypertension, and measures to reduce CypD acetylation can improve vascular function and reduce hypertension. Essential hypertension and animal models of hypertension are linked to inactivation of mitochondrial deacetylase Sirt3 by highly reactive lipid oxidation products, isolevuglandins (isoLGs), and supplementation of mice with mitochondria targeted scavenger of isoLGs, mito2HOBA, improves CypD deacetylation. To test the specific role of CypD-K166 acetylation, we developed CypD-K166R deacetylation mimic mutant mice. Mitochondrial respiration, vascular function and systolic blood pressure in CypD-K166R mice was similar to wild-type C57Bl/6J mice. Meanwhile, angiotensin II-induced hypertension was substantially attenuated in CypD-K166R mice (144 mmHg) compared with wild-type mice (161 mmHg). Angiotensin II infusion in wild-type mice significantly increased mitochondrial superoxide, impaired endothelial dependent relaxation, and reduced the level of endothelial nitric oxide which was prevented in angiotensin II-infused CypD-K166R mice. Hypertension is linked to increased levels of inflammatory cytokines TNFα and IL-17A promoting vascular oxidative stress and end-organ damage. We have tested if CypD-K166R mice are protected from cytokine-induced oxidative stress. Indeed, ex vivo incubation of aorta with the mixture of angiotensin II, TNFα and IL-17A (24 hours) increased mitochondrial superoxide by 2-fold in wild-type aortas which was abrogated in CypD-K166R mice. These data support the pathophysiological role of CypD acetylation in inflammation, oxidative stress and hypertensive end-organ damage. We propose that targeting CypD acetylation may have therapeutic potential in treatment of vascular dysfunction and hypertension.

Hypertension remains a major health problem in Western Societies, and blood pressure is poorly controlled in a third of patients despite use of multiple drugs. Mitochondrial dysfunction contributes to hypertension, and mitochondria-targeted agents can potentially improve treatment of hypertension. We have proposed that mitochondrial oxidative stress produces reactive dicarbonyl lipid peroxidation products, isolevuglandins, and that scavenging of mitochondrial isolevuglandins improves vascular function and reduces hypertension. To test this hypothesis, we have studied the accumulation of mitochondrial isolevuglandins-protein adducts in patients with essential hypertension and Ang II (angiotensin II) model of hypertension using mass spectrometry and Western blot analysis. The therapeutic potential of targeting mitochondrial isolevuglandins was tested by the novel mitochondria-targeted isolevuglandin scavenger, mito2HOBA. Mitochondrial isolevuglandins in arterioles from hypertensive patients were 250% greater than in arterioles from normotensive subjects, and ex vivo mito2HOBA treatment of arterioles from hypertensive subjects increased deacetylation of a key mitochondrial antioxidant, SOD2 (superoxide dismutase 2). In human aortic endothelial cells stimulated with Ang II plus TNF (tumor necrosis factor)-α, mito2HOBA reduced mitochondrial superoxide and cardiolipin oxidation, a specific marker of mitochondrial oxidative stress. In Ang II–infused mice, mito2HOBA diminished mitochondrial isolevuglandins-protein adducts, raised Sirt3 (sirtuin 3) mitochondrial deacetylase activity, reduced vascular superoxide, increased endothelial nitric oxide, improved endothelium-dependent relaxation, and attenuated hypertension. Mito2HOBA preserved mitochondrial respiration, protected ATP production, and reduced mitochondrial permeability pore opening in Ang II–infused mice. These data support the role of mitochondrial isolevuglandins in endothelial dysfunction and hypertension. We conclude that scavenging of mitochondrial isolevuglandins may have therapeutic potential in treatment of vascular dysfunction and hypertension.

Hypertension remains a major health problem in Western Societies, and blood pressure is poorly controlled in a third of patients despite use of multiple drugs. Mitochondrial dysfunction contributes to hypertension and mitochondria-targeted agents can potentially improve treatment of hypertension. We have proposed that mitochondrial oxidative stress produces reactive dicarbonyl lipid peroxidation products isolevuglandins (isoLGs) and that scavenging of mitochondrial isoLG improves vascular function and reduces hypertension. To test this hypothesis, we have studied the accumulation of mitochondrial isoLG-protein adducts in human patients with essential hypertension and angiotensin II mouse model of hypertension using mass spectrometry and Western blot analysis. The therapeutic potential of targeting mitochondrial isoLG was tested by the novel mitochondria-targeted isoLG scavenger, mito2HOBA. Mitochondrial isoLG in arterioles isolated from hypertensive patients were 250% greater than in arterioles from normotensive subjects, and ex vivo mito2HOBA treatment of arterioles from hypertensive subjects improved deacetylation of a key mitochondrial antioxidant, superoxide dismutase 2 (SOD2). In human aortic endothelial cells, mito2HOBA diminished mitochondrial superoxide and inhibited cardiolipin oxidation, a specific marker of mitochondrial oxidative stress. In angiotensin II-infused mice, mito2HOBA prevented accumulation of mitochondrial isoLG-protein adducts, improved Sirt3 mitochondrial deacetylase activity, reduced vascular superoxide, increased endothelial nitric oxide, improved endothelium-dependent relaxation, and attenuated hypertension. Mito2HOBA preserved mitochondrial respiration, protected ATP production, and reduced mitochondrial permeability pore opening in angiotensin II-infused mice. These data support the role of mitochondrial isoLGs in endothelial dysfunction and hypertension. We conclude that scavenging of mitochondrial isoLGs may have therapeutic potential in treatment of vascular dysfunction and hypertension.

Inflammation is a major cause of morbidity and mortality in Western societies. Despite use of multiple drugs, both chronic and acute inflammation still represent major health burdens. Inflammation produces highly reactive dicarbonyl lipid peroxidation products such as isolevuglandins which covalently modify and cross-link proteins via lysine residues. Mitochondrial dysfunction has been associated with inflammation; however, its molecular mechanisms and pathophysiological role are still obscure. We hypothesized that inflammation-induced isolevuglandins contribute to mitochondrial dysfunction and mortality. To test this hypothesis, we have (a) investigated the mitochondrial dysfunction in response to synthetic 15-E2-isolevuglandin (IsoLG) and its adducts; (b) developed a new mitochondria-targeted scavenger of isolevuglandins by conjugating 2-hydroxybenzylamine to the lipophilic cation triphenylphosphonium, (4-(4-aminomethyl)-3-hydroxyphenoxy)butyl)-triphenylphosphonium (mito2HOBA); (c) tested if mito2HOBA protects from mitochondrial dysfunction and mortality using a lipopolysaccharide model of inflammation. Acute exposure to either IsoLG or IsoLG adducts with lysine, ethanolamine or phosphatidylethanolamine inhibits mitochondrial respiration and attenuates Complex I activity. Complex II function was much more resistant to IsoLG. We confirmed that mito2HOBA markedly accumulates in isolated mitochondria and it is highly reactive with IsoLGs. To test the role of mitochondrial IsoLGs, we studied the therapeutic potential of mito2HOBA in lipopolysaccharide mouse model of sepsis. Mito2HOBA supplementation in drinking water (0.1 g/L) to lipopolysaccharide treated mice increased survival by 3-fold, improved complex I-mediated respiration, and histopathological analyses supported mito2HOBA-mediated protection of renal cortex from cell injury. These data support the role of mitochondrial IsoLG in mitochondrial dysfunction and inflammation. We conclude that reducing mitochondrial IsoLGs may be a promising therapeutic target in inflammation and conditions associated with mitochondrial oxidative stress and dysfunction., Graphical abstract Image 1, Highlights • Isolevuglandins and their adducts inhibit mitochondrial respiration and attenuates Complex I activity. • 2-Hydroxybenzylamine conjugated to triphenylphosphonium, mito2HOBA, accumulates in mitochondria. • Mito2HOBA in drinking water improves complex I-mediated respiration in LPS model of sepsis. • Mitochondria-targeted scavenger of isolevuglandins mito2HOBA reduces mortality in LPS model.

One third of adult population has hypertension and third of hypertensive patients remain hypertensive despite treatment with multiple drugs. There have been no mechanistically novel treatments for hypertension in the past 30 years. We propose that new agents targeting CypD activation, a regulatory subunit of mitochondrial permeability transition pore, could add to the currently available therapeutic armamentarium to improve treatment of hypertension. To test this hypothesis we have studied CypD acetylation which was associated with CypD activation. We found that angiotensin II-induced hypertension is linked to CypD-K166 hyperacetylation, mPTP opening, impaired mitochondrial respiration and reduced kidney ATP. CypD hyperacetylation may result from imbalance between GCN5L1 acetyltransferase and reduced Sirt3 deacetylase activity. Indeed, aortic mitochondrial level of GCN5L1 increased by 50% while Sirt3 was reduced by 40% in angiotensin II infused mice. Sirt3 can be inactivated by cytotoxic reactive lipid dicarbonyls derived from arachidonic acid, isoketals, leading to mitochondrial hyperacetylation. To test the role of mitochondrial isoketals we synthesized a novel mitochondria-targeted isoketal scavenger mito2HOBA. mito2HOBA supplementation in drinking water (0.1g/Liter) attenuates angiotensin II-induced hypertension, prevents accumulation of mitochondrial isoketal protein adducts in the aorta and kidney measured by LC/MS and D11 antibody, improves Sirt3/GCNL1 ratio, normalizes the CypD acetylation, attenuates mPTP opening, protects mitochondrial respiration and preserves normal kidney ATP level. Decreased NO bioavailability is a hallmark of endothelial oxidative stress in hypertension due to superoxide overproduction and NO oxidation. Interestingly, angiotensin II increases vascular superoxide by 2-fold and reduces endothelial NO level to 50% while mito2HOBA diminishes vascular O 2 • and preserves NO bioavailability in angiotensin II-infused mice. These data support the role of isoketals-mediated CypD hyperacetylation in endothelial dysfunction and hypertension. We conclude that scavenging of mitochondrial isoketals may have therapeutic potential in treatment of hypertension and target-organ-damage.

This paper is devoted to morphological tagging task for Russian. There are multiple corpora created for the task during the years of research. Unfortunately, these corpora often have incompatible annotation guidelines and tag sets. This makes it difficult to utilize more than one corpus for training machine learning methods. Several attempts to manually unify guidelines and tag sets took place in the past. The most recent one was MorphoRuEval-2017 evaluation where organizers decided to unify annotations of four corpora. In this paper we propose a morphological tagging method which is able to be trained on any set of corpora with arbitrary guidelines and tag sets without any manual mapping provided. The method is based on transfer learning technique successfully applied in multilingual and cross-domain research. Firstly we establish a couple of solid neural network-based morphological tagging baselines. They both employ bidirectional LSTM networks but utilize different word embeddings. Comparative study on multiple corpora shows that fasttext model clearly outperforms word2vec one. Further we extend these baselines with transfer learning application. We prove transfer learning-based method effectiveness by performing two series of experiments. In the first series we exploit three wellknown corpora with incompatible tag sets: RNC, SynTagRus (original and Universal Dependencies versions). In the second series we utilize four corpora from MorphoRuEval-2017 evaluation with tag sets preliminary unified by organizers. In both cases transfer learning application improves performance of the method while training on more corpora. In the second case it even surpasses traditional joint training.

Mitochondria are the major source of superoxide radicals and superoxide overproduction contributes to cardiovascular diseases and metabolic disorders. Endothelial dysfunction and diminished nitric oxide levels are early steps in the development of these pathological conditions. It is known that physiological production of nitric oxide reduces oxidative stress and inflammation, however, the precise mechanism of “antioxidant” effect of nitric oxide is not clear. In this work we tested the hypothesis that physiological levels of nitric oxide diminish mitochondrial superoxide production without inhibition of mitochondrial respiration. In order to test this hypothesis we analyzed effect of low physiological fluxes of nitric oxide (20 nM/min) on superoxide and hydrogen peroxide production by ESR spin probes and Amplex Red in isolated rat brain mitochondria. Indeed, low levels of nitric oxide substantially attenuated both basal and antimycin A-stimulated production of reactive oxygen species in the presence of succinate or glutamate/malate as mitochondrial substrates. Furthermore, slow releasing NO donor DPTA-NONOate (100 μM) did not change oxygen consumption in State 4 and State 3. However, the NO-donor strongly inhibited oxygen consumption in the presence of uncoupling agent CCCP, which is likely associated with inhibition of the over-reduced complex IV in uncoupled mitochondria. We have examined accumulation of dinitrosyl iron complexes and nitrosothiols in mitochondria treated with fast-releasing NO donor MAHMA NONOate (10 μM) for 30 min until complete release of NO. Following treatment with NO donor, mitochondria were frozen for direct detection of dinitrosyl iron complexes using Electron Spin Resonance (ESR) while accumulation of nitrosothiols was measured by ferrous-N-Methyl-D-glucamine dithiocarbamate complex, Fe(MGD)2, in lysed mitochondria. Treatment of mitochondria with NO-donor gave rise to ESR signal of dinitrosyl iron complexes while ESR spectra of Fe(MGD)2 supplemented mitochondrial lysates showed presence of both dinitrosyl iron complexes and nitrosothiols. We suggest that nitric oxide attenuates production of mitochondrial superoxide by post-translational modifications by nitrosylation of protein cysteine residues and formation of protein dinitrosyl iron complexes with thiol-containing ligands and, therefore, nitric oxide reduction in pathological conditions associated with endothelial dysfunction may increase mitochondrial oxidative stress.

A framework for fast text analysis, which is developed as a part of the Texterra project, is described. Texterra provides a scalable solution for the fast text processing on the basis of novel methods that exploit knowledge extracted from the Web and text documents. For the developed tools, details of the project, use cases, and evaluation results are presented.

It has been established previously that up to 40% of mouse CD34(+) hematopoietic stem cells are capable of internalizing exogenous dsDNA fragments both in vivo and ex vivo. Importantly, when mice are treated with a combination of cyclophosphamide and dsDNA, the repair of interstrand crosslinks in hematopoietic progenitors is attenuated, and their pluripotency is altered. Here we show for the first time that among various actively proliferating mammalian cell populations there are subpopulations capable of internalizing dsDNA fragments. In the context of cancer, such dsDNA-internalizing cell subpopulations display cancer stem cell-like phenotype. Furthermore, using Krebs-2 ascites cells as a model, we found that upon combined treatment with cyclophosphamide and dsDNA, engrafted material loses its tumor-initiating properties which we attribute to the elimination of tumor-initiating stem cell subpopulation or loss of its tumorigenic potential.

The paper presents a framework for fast text analytics developed during the Texterra project. Texterra delivers a scalable solution for text processing based on novel methods that exploit knowledge extracted from the Web and text documents. This paper describes details of the project, use-cases and results of evaluation for all developed tools.

Horizontal transfer (HT) is a complex phenomenon usually used as an explanation of phylogenetic inconsistence, which cannot be interpreted in terms of vertical evolution. Most examples of HT of eukaryotic genes involve transposable elements. An intriguing feature of HT is that its frequency differs among transposable elements classes. Although HT is well known for DNA transposons and long terminal repeat (LTR) retrotransposons, non-LTR retrotransposons rarely undergo HT, and their phylogenies are largely congruent to those of their hosts. Previously, we described HT of CR1-like non-LTR retrotransposons between butterflies (Maculinea) and moths (Bombyx), which occurred less than 5 million years ago (Novikova O, Sliwinska E, Fet V, Settele J, Blinov A, Woyciechowski M. 2007. CR1 clade of non-LTR retrotransposons from Maculinea butterflies (Lepidoptera: Lycaenidae): evidence for recent horizontal transmission. BMC Evol Biol. 7:93). In this study, we continued to explore the diversity of CR1 non-LTR retrotransposons among lepidopterans providing additional evidences to support HT hypothesis. We also hypothesized that DNA transposons could be involved in HT of non-LTR retrotransposons. Thus, we performed analysis of one of the groups of DNA transposons, mariner-like DNA elements, as potential vectors for HT of non-LTR retrotransposons. Our results demonstrate multiple HTs between Maculinea and Bombyx genera. Although we did not find strong evidence for our hypothesis of the involvement of DNA transposons in HT of non-LTR retrotransposons, we demonstrated that recurrent and/or simultaneous flow of TEs took place between distantly related moths and butterflies.

Enzymatic deamination of bases in DNA or RNA leads to an alteration of flow of genetic information. Adenosine deaminases edit RNA (ADARs, TADs). Specialized cytidine deaminases are involved in RNA/DNA editing in lipid metabolism (APOBEC1) and in innate (APOBEC3 family) and humoral (AID) immunity. APOBEC2 is required for proper muscle development and, along with AID, was implicated in demethylation of DNA. The functions of APOBEC4, APOBEC5, and other deaminases recently discovered by bioinformatics approaches are unknown. What is the basis for the diverse biological functions of enzymes with similar enzyme structure and the same principal enzymatic reaction? AID, APOBEC1, lamprey CDA1, and APOBEC3G enzymes cause uracil DNA glycosylase-dependent induction of mutations when overproduced ectopically in bacteria or yeast. APOBEC2, on the contrary, is nonmutagenic. We studied the effects of the expression of various deaminases in yeast and bacteria. The mutagenic specificities of four deaminases, hAID, rAPOBEC1, hAPOBEC3G, and lamprey CDA1, are strikingly different. This suggests the existence of an intrinsic component of deaminase targeting. The expression of yeast CDD1 and TAD2/TAD3, human APOBEC4, Xanthomonas oryzae APOBEC5, and deaminase encoded by Micromonas sp. gene MICPUN_56782 was nonmutagenic. A lack of a mutagenic effect for Cdd1 is expected because the enzyme functions in the salvage of pyrimidine nucleotides, and it is evolutionarily distant from RNA/DNA editing enzymes. The reason for inactivity of deaminases grouped with APOBEC2 is not obvious from their structures. This can not be explained by protein insolubility and peculiarities of cellular distribution and requires further investigation.

The genus Triticum L. includes the major cereal crop, common or bread wheat (hexaploid Triticum aestivum L.), and other important cultivated species. Here, we conducted a phylogenetic analysis of all known wheat species and the closely related Aegilops species. This analysis was based on chloroplast matK gene comparison along with trnL intron sequences of some species. Polyploid wheat species are successfully divided only into two groups – Emmer (sections Dicoccoides and Triticum) and Timopheevii (section Timopheevii). Results reveal strictly maternal plastid inheritance of synthetic wheat amphiploids included in the study. A concordance of chloroplast origin with the definite nuclear genomes of polyploid species that were inherited at the last hybridization events was found. Our analysis suggests that there were two ancestral representatives of Aegilops speltoides Tausch that participated in the speciation of polyploid wheats with B and G genome in their genome composition. However, G genome species are younger in evolution than ones with B genome. B genome-specific PCR primers were developed for amplification of Acc-1 gene.

Purpose: Autosomal dominant optic atrophy is a form of blindness, due in part to mutations affecting the mitochondrial-targeted OPA1 gene product. Both OPA1-positive and OPA1-negative families exhibit variable expressivity and incomplete penetrance. The purpose of this study was therefore to determine if the background mtDNA genotype acts as a genetic modifier for the expression of this disease. Methods: To find novel pathogenic OPA1 mutations, we performed complete OPA1 gene exon sequencing in 30 patients. To assess the possibility that mitochondrial DNA haplotype acts as a genetic modifier, we determined the mitochondrial DNA haplotype in 29 Caucasian OPA1-positive and OPA1-negative patients. Deviations in haplotype distribution between patient and control groups were determined by statistical means. Results: Seven new pathogenic OPA1 mutations were found. Most were detected in the mitochondrial targeting N-terminus or in the coiled-coil domain at the C-terminus. Mitochondrial DNA haplotype analysis indicated that the European haplogroup distribution was different between Caucasian patients and controls. Further, haplogroup J was three-fold over-represented in OPA1-negative patients. Conclusions: Overall, our results support haploinsufficiency as a genetic mechanism in OPA1-positive cases and also suggest that mtDNA genetic background may influence disease expression in a subset of cases.

Molecular phylogenetic methods were used to examine morphologically based hypotheses concerning the taxonomic structure and relationships of the grasshopper subfamily Gomphocerinae. Two mitochondrial gene (cytochrome b and cytochrome oxidase subunit I) sequences were determined for twenty-five species representing eleven Palaearctic genera. The studied Gomphocerinae species constituted a monophyletic group; furthermore, the earlier division of Gomphocerinae into tribes was supported, with each tribe monophyletic. There was no support for various systems uniting Stenobothrini and Gomphocerini into one tribe. Two separate clusters were discerned in Gomphocerini and two tribes were distinguished – Gomphocerini (genera Aeropus, Stauroderus, Chorthippus) and Stenobothrini (genera Omocestus, Stenobothrus).

A full-length copy of the F6.2 gene from the tissue-specific BRa locus of the Chironomus thummi chromosome IV was isolated and analyzed. The gene contains two exons (715 and 644 bp, respectively) and one 172-bp intron. The data of the RT-PCR analysis demonstrated that F6.2 was transcriptionally active at different developmental stages of Chironomus thummi and at least in the last larval stage of C. dorsalis. The distribution of theF6.2gene among 42 species ofChironomus, as well as among two other genera of the family Chironomidae was examined by means of PCR. The F6.2 sequence was found in 34 Chironomus species. Using in situ hybridization, three species were analyzed for the presence of the F6.2 homologous sequences. In five species, the sequence of the F6.2PCR product was determined. In these species, the intron size polymorphism caused by the variation of the number of the intron-forming repeats was observed. The data obtained provided evaluation of the F6.2 distribution among the genus Chironomus.

The presence of repetitive elements can create serious problems for sequence analysis, especially in the case of homology searches in nucleotide sequence databases. Repetitive elements should be treated carefully by using special programs and databases. In this paper, various aspects of SINE (short interspersed repetitive element) identification, analysis and evolution are discussed.

Several LINE-1s have been isolated and characterized from genomic DNA of the vole, Microtus kirgisorum. Blot hybridization revealed specific restriction patterns of L1 elements in vole genomes. Rehybridization of the genomic blot with a cloned 5'-end fragment revealed two major bands indicating the presence of two different L1 subfamilies. The copy numbers are estimated for different parts of M. kirgisorum L1 elements. Data also demonstrate that most vole L1 elements are truncated at the 5'-end; however, in contrast to mouse, the ORF1 copy number is higher in vole. A difference between the substitution rates of the ORF1 5'-region (approximately 330 nucleotides) and the rest of the L1 coding regions is revealed.

A repetitive DNA sequence, MS2, was isolated from EcoRI-digested genomic DNA of the vole, Microtus subarvalis. The fragment was cloned and sequenced. Sequence analysis of this 1194-bp fragment revealed a 156-bp region demonstrating a 55% homology with the mouse B1 repeat. The remaining MS2 sequence shows no significant homology with other known Gen-Bank sequences. The results of in situ hybridization of MS2 on vole metaphase chromosomes indicate the fragment is confined to heterochromatin blocks of the sex chromosomes in all but one species (M. arvalis). Distribution of MS2 sequences provides evidence for heterogeneity of the giant heterochromatin blocks of the XY Chromosomes (Chrs) in voles, for the unique cluster-like localization of MS2 within these blocks.

Free radical oxidation of arachidonic acid produces highly reactive isolevuglandins (isoLG) which causes mitochondrial dysfunction by opening of the mitochondrial permeability transition pore (mPTP). Inhibition of the mPTP regulatory subunit cyclophilin D reduces isoLG-induced mitochondrial dysfunction (Free Radic Biol Med 2010;49:567-79). We found that depletion of cyclophilin D diminishes mitochondrial O 2 ●– , improves vascular relaxation and reduces hypertension (Hypertension 2016;67:1218-27). We hypothesized that hypertension is associated with accumulation of mitochondrial isoLG and mitochondria-targeted isoLG scavenger reduces vascular oxidative stress and attenuates hypertension. To test this hypothesis we developed mitochondria-targeted isoLG scavenger 4-(aminomethyl)-3-hydroxyphenoxy) butyl)triphenyl-phosphonium (mito2HOBA) by conjugation of lipophilic triphenylmethylphosphonium cation to 2-hydroxybenzylamine (2HOBA). Mito2HOBA is a water soluble compound which was well tolerated by cultured human aortic endothelial cells (HAECs) and by mice receiving it in drinking water. It was found that mito2HOBA (50 nM) inhibited mitochondrial O 2 ●– production (MitoSOX/HPLC) and prevented cardiolipin oxidation (LS-MS) in HEACs incubated with TNFα+Angiotensin II while untargeted analog 2HOBA (50 nM) was not effective. The functional role of mitochondrial isoLG was tested in vivo using angiotensin II model of hypertension. C57Bl/6J mice were infused with angiotensin II (0.7 mg/kg/day) or saline (Sham) and received mito2HOBA in the drinking water (0.1 g/L). It was found that mito2HOBA diminished angiotensin II-induced hypertension. Hypertension was associated with increased formation of mitochondrial isoLG measured by Western blot analysis of heart mitochondria using the anti-isoLG D11 scfv antibody and 2-fold decrease in kidney ATP. mito2HOBA reduced accumulation of isoLG adducts in the heart mitochondria and prevented drop in kidney ATP in angiotensin II-infused mice. Decrease of NO is a hallmark of endothelial dysfunction in hypertension due to vascular oxidative stress. We examined vascular oxidative stress by measurements of aortic O 2 ●– using fluorescent O 2 ●– probe DHE and HPLC, and analysis of endothelial NO using Electron Spin Resonance and specific NO spin trap Fe(DETC) 2 . It was found that mito2HOBA reduced vascular O 2 ●– in angiotensin II-infused mice and preserved endothelial NO. These data support the role of mitochondrial oxidative stress and mitochondrial isoLG in endothelial dysfunction and hypertension. We conclude that scavenging of mitochondrial isoLG may have therapeutic potential in cardiovascular disease.

–derived SINE, specific forthree rodent families: Muridae, Cricetidae, and Spalacidae. Theseelements are composed of three regions: a 58 RNA-related region(RNA-polymerase III promoter), a tRNA-unrelated region, and a38 AT-rich region (polyadenylation signals); the total length of aB2 element is approximately 200 bp (Deininger 1989; Weiner etal. 1986; Rogers 1985; Krayev et al. 1982).Previously, Kass et al. (1997) and Serdobova and Kramerov(1998) concluded B2 elements do not exist in the human genome.Our computer analysis of sequences from nucleotide databases(Altschul et al. 1990; Benson et al. 1999) revealed several humansequences with significant homology to the rodent B2 consensussequence (Bains and Temple-Smith 1989; Jurka et al. 1992; Okadaand Oshima 1995). However, some of these sequences were ob-tained from human–rodent hybrid cells and may represent con-tamination or transposition within the cells of the hybrids. Toverify the presence of B2 SINEs in the human genome, we de-signed oligonucleotide primers from the mouse B2 element con-sensus sequence. The 58 primer identified the tRNA-related region,and the 38 primer identified the 38 region immediately upstreamfrom the AT-rich sequences of the B2 consensus sequence (Fig. 1).Human genomic DNA from different sources (placenta, blood,and commercial placental DNA) was amplified, cloned, and se-quenced. The average divergence of these sequences from therodent B2 consensus sequences (Kass et al. 1997) is about 6%.However, only one specific subclass of human B2 elements, am-plified with mouse B2 primer sequences, was analyzed, and con-clusions about the divergence of all human B2 elements cannot beattempted. Examples of potential human B2 elements from sub-telomeric DNA sequences (Ning and Flint 1996) with significantly

Background Autosomal dominant optic atrophy (ADOA), a form of progressive bilateral blindness due to loss of retinal ganglion cells and optic nerve deterioration, arises predominantly from mutations in the nuclear gene for the mitochondrial GTPase, OPA1. OPA1 localizes to mitochondrial cristae in the inner membrane where electron transport chain complexes are enriched. While OPA1 has been characterized for its role in mitochondrial cristae structure and organelle fusion, possible effects of OPA1 on mitochondrial function have not been determined. Results Mitochondria from six ADOA patients bearing OPA1 mutations and ten ADOA patients with unidentified gene mutations were studied for respiratory capacity and electron transport complex function. Results suggest that the nuclear DNA mutations that give rise to ADOA in our patient population do not alter mitochondrial electron transport. Conclusion We conclude that the pathophysiology of ADOA likely stems from the role of OPA1 in mitochondrial structure or fusion and not from OPA1 support of oxidative phosphorylation.

Retrotransposons are the major component of plant genomes. Chromodomain-containing Gypsy long terminal repeat (LTR) retrotransposons are widely distributed in eukaryotes. Four distinct clades of chromodomain-containing Gypsy retroelements are known from the vascular plants: Reina, CRM, Galadriel and Tekay. At the same time, almost nothing is known about the repertoire of LTR retrotransposons in bryophyte genomes. We have combined a search of chromodomain-containing Gypsy retroelements in Physcomitrella genomic sequences and an experimental investigation of diverse moss species. The computer-based mining of the chromodomain-containing LTR retrotransposons allowed us to describe four different elements from Physcomitrella. Four novel clades were identified that are evolutionarily distinct from the chromodomain-containing Gypsy LTR retrotransposons of other plants.

Leber's hereditary optic neuropathy (LHON) causes central vision loss from bilateral optic neuropathy. Although 13 mitochondrial DNA (mtDNA) mutations are strongly associated with LHON, only three account for roughly 90% of cases and thus are found in multiple independent LHON families. The remaining LHON mutations are rare. Here, we describe the clinical and genetic characterization of a new LHON mtDNA mutation. The 12848T mutation alters a highly conserved amino acid in the ND5 complex I gene, is not found in controls, and is heteroplasmic. Despite ND5 being the largest of the mtDNA complex I genes, ND5 mutations are quite rare in LHON.

DNA polymerase (pol) η participates in hypermutation of A:T bases in Ig genes because humans deficient for the polymerase have fewer substitutions of these bases. To determine whether polymerase η is also responsible for the well-known preference for mutations of A vs T on the nontranscribed strand, we sequenced variable regions from three patients with xeroderma pigmentosum variant (XP-V) disease, who lack polymerase η. The frequency of mutations in the intronic region downstream of rearranged JH4 gene segments was similar between XP-V and control clones; however, there were fewer mutations of A:T bases and correspondingly more substitutions of C:G bases in the XP-V clones (p < 10−7). There was significantly less of a bias for mutations of A compared with T nucleotides in the XP-V clones compared with control clones, whereas the frequencies for mutations of C and G were identical in both groups. An analysis of mutations in the WA sequence motif suggests that polymerase η generates more mutations of A than T on the nontranscribed strand. This in vivo data from polymerase η-deficient B cells correlates well with the in vitro specificity of the enzyme. Because polymerase η inserts more mutations opposite template T than template A, it would generate more substitutions of A on the newly synthesized strand.

Highly repetitive DNA sequences of the LINE-1 (L1) family, which are interspersed in the genomes of various organisms, are among the many transposable elements deserving attention. L1 elements are widely distributed throughout the mammalian genomes (Burton et al. 1986). Their occurrence was first demonstrated in man and other mammals. Subsequently, L1like elements were found to occur in insects and higher plants (DiNosera and Sakaki 1990). No definite role has yet been assigned to L1 elements. Their presence in virtually all organisms suggests that they might be important for the structural organization and function of the genome. The sequence features of L 1 include two open reading frames (ORFs), the short 5' proximal ORF1 and the about four times longer 3' proximal ORF2. L1 element contains a poly(A) tail at the 3' end and is flanked by short direct repeats (Fanning and Singer 1987). The sequence similarity between L1 and retroviruses and related transposable elements suggests that the predicted ORF2 encoded protein is involved in the transpos i t ion of L1 e lement . The suppor t for the involvement of LI-encoded proteins in L1 transposition has come from in vitro studies (Leibold et al. 1990). More recently it was shown that human L1 element encodes reverse transcriptase activity (Dombroski et al. 1991; Mathias et al. 1991). We describe here genomic L1 element detected in voles of the Microtus genus. Genomic DNA was prepared from vole liver according to standard procedures (Henry et al. 1990); complete digestion was performed with endonuclease EcoRI. Comparative restriction analysis of genomic DNAs from five vole species is shown in Fig. 1. Clearly, the number and distribution of bands corresponding to DNA repeats are specific to each of the species. The intense band is common to the four species, and it represents the same

Hereditary neuropathy with liability to pressure palsies (HNPP) is a recurrent disorder of the peripheral nervous system characterized by reversible episodes of sensorimotor deficits after neural compression injuries. Also known as tomaculous neuropathy, HNPP is further characterized ultrastructurally by multiple focal thickenings (tomacula) of peripheral myelin and has an autosomal dominant inheritance. The neuropathology of HNPP includes a partial deletion encoding the peripheral myelin protein 22 (PMP-22) gene on chromosome 17, resulting in underexpression of PMP-22. We describe multiple compression mononeuropathies in an individual with HNPP and report neuropathologic findings in 2 clinically asymptomatic family members. Diagnosis was confirmed using pulsed-field gel electrophoresis. We believe that this diagnosis is clinically underappreciated by hand surgeons and should be considered in the differential diagnosis of patients with atypical presentations of compression neuropathies.

Background Antibody genes are diversified by somatic hypermutation (SHM), gene conversion and class-switch recombination. All three processes are initiated by the activation-induced deaminase (AID). According to a DNA deamination model of SHM, AID converts cytosine to uracil in DNA sequences. The initial deamination of cytosine leads to mutation and recombination in pathways involving replication, DNA mismatch repair and possibly base excision repair. The DNA sequence context of mutation hotspots at G-C pairs during SHM is DGYW/WRCH (G-C is a hotspot position, R = A/G, Y = T/C, W = A/T, D = A/G/T). Results To investigate the mechanisms of AID-induced mutagenesis in a model system, we studied the genetic consequences of AID expression in yeast. We constructed a yeast vector with an artificially synthesized human AID gene insert using codons common to highly expressed yeast genes. We found that expression of the artificial hAIDSc gene was moderately mutagenic in a wild-type strain and highly mutagenic in an ung1 uracil-DNA glycosylase-deficient strain. A majority of mutations were at G-C pairs. In the ung1 strain, C-G to T-A transitions were found almost exclusively, while a mixture of transitions with 12% transversions was characteristic in the wild-type strain. In the ung1 strain mutations that could have originated from deamination of the transcribed stand were found more frequently. In the wild-type strain, the strand bias was reversed. DGYW/WRCH motifs were preferential sites of mutations. Conclusion The results are consistent with the hypothesis that AID-mediated deamination of DNA is a major cause of mutations at G-C base pairs in immunoglobulin genes during SHM. The sequence contexts of mutations in yeast induced by AID and those of somatic mutations at G-C pairs in immunoglobulin genes are significantly similar. This indicates that the intrinsic substrate specificity of AID itself is a primary determinant of mutational hotspots at G-C base pairs during SHM.