[Dynamics of sperm indicators in men with chronic prostatitis associated with secondary infertility based on the use of ANDROEXPERT SV1 suppositories].

INTRODUCTION: Chronic prostatitis (CP) is the most common cause of a disorder of not only sexual, but also reproductive functions in men, which is caused by a decrease in the quality of the ejaculate and is confirmed by a deterioration in spermogram parameters, up to oligoasthenoteratozoospermia and azoospermia. In such a situation, the onset of pregnancy in the natural cycle is impossible - even with a completely preserved reproductive function in a woman. As an active substance in the recovery of men with chronic prostatitis associated with secondary infertility, we proposed oxidised dextran in the form of a pharmaceutical composition, which was developed at Federal Research Centre for Fundamental and Translational Medicine, Novosibirsk, Russia; and produced by ANDROEXPERT Research and Manufacturing Company LTD, Novosibirsk, Russia. The purpose of the study was to evaluate the dynamics of spermogram parameters as a result of the use of suppositories with oxidised dextran "ANDROEXPERT SV1" in men with chronic prostatitis associated with secondary infertility. MATERIAL AND METHODS: The study included 52 patients with chronic prostatitis associated with secondary infertility. The mean age of the patients was 32+/-2.2 years. The examination of patients was carried out in accordance with the algorithm recommended by the Russian Society of Urology, the European Urological Association and the standard of primary health care for chronic prostatitis (Appendix to the order of the Ministry of Health of Russia No. 775n of 09.11.2012). The patients were divided into two groups: the 1st group of patients received oxidised dextran rectally once per day for 10 days, then 20 days every other day; The 2nd (comparison group) received a placebo in the form of rectal suppositories consisting of cocoa butter in a similar way. RESULTS: The volume of ejaculate among patients of the 1st group increased from 2.54+/-0.15 to 3.13+/-0.18 (ml), which amounted to 23.23%; p<0.05. The sperm concentration indicator increased from 26.40+/-4.18 to 41.12+/-5.37 (million/ml) - by 55.75%; p<0.05, which led to an increase in the total number of spermatozoa in the ejaculate by 88.5%, from 65.47+/-8.41 to 123.41+/-12.33 (million pieces); p<0.05. The number of leukocytes in the same group decreased by 2.8 times: from 1.25+/-0.17 to 0.44+/-0.10 (million/ml) - by 64.80%; p<0.05. The indicator of the severity of sperm agglutination decreased by 3 times: from 0.52+/-0.07 to 0.17+/-0.03 - by 0.35 arb. units, which was 67.31%; p<0.05. The indicators of the spermogram in patients with chronic prostatitis of the second group did not change in the course of treatment. CONCLUSION: Assessing the dynamics of spermogram parameters as a result of the use of "ANDROEXPERT SV1" in men with chronic prostatitis associated with secondary infertility, there is a decrease in the number of leukocytes and a significant improvement in the parameters of the ejaculate, both its general properties and the main characteristics of spermatozoa. Thus, ANDROEXPERT SV1 suppositories show high clinical efficacy and safety in monotherapy and may be promising when choosing the management tactics for this group of patients.

[The use of rectal suppositories with oxidised dextran for calculous prostatitis].

This article presents its own experience of using suppositories with oxidised dextran in the treatment of a patient with recurrent bacterial prostatitis complicated by prostate calcifications, these are the positive effects on the use of suppositories with oxidised dextran: a decrease in the size of calcified areas, a stable elimination of opportunistic microflora and the achievement of long-term remission of the disease.

[Experimental study on the efficiency of rectal suppositories with oxidized dextran in patients with benign prostatic hyperplasia].

AIM: to evaluate therapeutic activity of oxidized dextran after rectal administration in a modified sulpiride model of benign prostatic hyperplasia in mice. MATERIALS AND METHODS: The therapeutic activity of oxidized dextran was studied in an experimental model of benign prostatic hyperplasia in mice (sulpiride model). All in vivo studies were carried out on 30 male BALB/c mice with an average body weight of 30-35 g. RESULTS: It was found that rectal administration of a solution of oxidized dextran effectively neutralizes pathomorphological changes in experimental model of benign prostatic hyperplasia. In the group of animals, which received 2% solution of oxidized dextran as treatment, the most prominent therapeutic effect in an experimental model was seen and it was confirmed by a decrease in the bladder detrusor thickness and the normalization of its lumen.

Photosynthetic Picoeukaryotes in the Land-Fast Ice of the White Sea, Russia.

The White Sea is a unique marine environment combining features of temperate and Arctic seas. The composition and abundance of photosynthetic picoeukaryotes (PPEs) were investigated in the land-fast ice of the White Sea, Russia, in March 2013 and 2014. High-throughput tag sequencing (Illumina MiSeq system) of the V4 region of the 18S rRNA gene was used to reveal the diversity of PPE ice community. The integrated PPE abundance varied from 11 × 106 cells/m2 to 364 × 106 cells/m2; the integrated biomass ranged from 0.02 to 0.26 mg С/m2. The composition of sea-ice PPEs was represented by 16 algae genera belonging to eight classes and three super-groups. Chlorophyta, especially Mamiellophyceae, dominated among ice PPEs. The detailed analysis revealed the latent diversity of Micromonas and Mantоniella. Micromonas clade E2 revealed in the subarctic White Sea ice indicates that the area of distribution of this species is wider than previously thought. We suppose there exists a new Micromonas clade F. Micromonas clade C and Minutocellulus polymorphus were first discovered in the ice and extend the modern concept of sympagic communities' diversity generally and highlights the importance of further targeting subarctic sea ice for microbial study.

The Mitochondrial Genome of the Moss Brachythecium rivulare (Hypnales, Brachytheciaceae).

The mitochondrial genome of the pleurocarpous moss Brachythecium rivulare has been sequenced and annotated. The genome consists of 104,460 base pairs and has approximately the same gene set and organization as other bryophyte mitogenomes. Whole mitochondrial genome comparison between B. rivulare and Physcomitrella patens, Tetraphis pellucida, Anomodon rugelii, and Anomodon attenuatus was performed. The primary cause of bryophyte mitochondrial gene length variation was found to be numerous indels in the introns. Bryophyte mitochondrial gene conservation level was estimated, and it was in a good congruence with the overall phylogeny of bryophytes with the percentage of mitogenome similarity being proportional to the age estimated by phylochronologic analysis. Annotation discrepancies in the analyzed mitogenome sequences were identified. The simple sequence repeat (SSR) content was evaluated, and candidate sites of RNA editing were predicted in the B. rivulare mitochondrial genome.

Metagenomics of Bolidophyceae in Plankton and Ice of the White Sea.

The molecular diversity of poorly studied algae of Bolidophyceae class was first estimated by Illumina sequencing of V4 region of 18S rRNA gene in ice, under-ice water and summer water of the subarctic White Sea. We used two clustering thresholds - 93 and 97% - and revealed 31 phylotypes of Bolidophyceae. Triparma pacifica and T. strigata were identified to species level. The association of individual phylotypes to certain biotopes (ice or plankton) and stages of seasonal succession (under ice or summer plankton) has been established. Some phylotypes are found in different biotopes and over a wide temperature range. Due to changing their genetic composition, Bolidophyceae are a constant component of the photoautotrophic plankton and ice communities.

Structure of Intergenic Spacer IGS1 of Ribosomal Operon from Schistidium Mosses.

The structure of the intergenic spacer 1 (IGS1) of the ribosomal operon from 12 species of Schistidium mosses was studied. In the IGS1 sequences of these species, three conserved regions and two areas of GC- and A-enriched repeats were identified. All of the studied mosses have a conserved pyrimidine-enriched motif at the 5'-end of IGS1. Species-specific nucleotide substitutions and insertions were found in the conserved areas. The repeated units contain single nucleotide substitutions that make unique the majority of repeated units. The positions of such repeats in IGS1 are species-specific, but their number can vary within the species and among operons of the same specimen. The comparison of IGS1 sequences from the Schistidium species and from representatives of ten other moss genera revealed the presence of common conserved motifs with similar localization. Presumably, these motifs are elements of termination of the pre-rRNA transcription and processing of rRNA.

Moss Phylogeny Reconstruction Using Nucleotide Pangenome of Complete Mitogenome Sequences.

Stability of composition and sequence of genes was shown earlier in 13 mitochondrial genomes of mosses (Rensing, S. A., et al. (2008) Science, 319, 64-69). It is of interest to study the evolution of mitochondrial genomes not only at the gene level, but also on the level of nucleotide sequences. To do this, we have constructed a "nucleotide pangenome" for mitochondrial genomes of 24 moss species. The nucleotide pangenome is a set of aligned nucleotide sequences of orthologous genome fragments covering the totality of all genomes. The nucleotide pangenome was constructed using specially developed new software, NPG-explorer (NPGe). The stable part of the mitochondrial genome (232 stable blocks) is shown to be, on average, 45% of its length. In the joint alignment of stable blocks, 82% of positions are conserved. The phylogenetic tree constructed with the NPGe program is in good correlation with other phylogenetic reconstructions. With the NPGe program, 30 blocks have been identified with repeats no shorter than 50 bp. The maximal length of a block with repeats is 140 bp. Duplications in the mitochondrial genomes of mosses are rare. On average, the genome contains about 500 bp in large duplications. The total length of insertions and deletions was determined in each genome. The losses and gains of DNA regions are rather active in mitochondrial genomes of mosses, and such rearrangements presumably can be used as additional markers in the reconstruction of phylogeny.

Metagenomic Analyses of White Sea Picoalgae: First Data.

Picoalgae (defined as cells smaller than 2-3 µm) include members of diverse taxonomic groups. They are an important constituent of marine plankton and ice biota and play a significant ecological role in biogeochemical cycles. Despite their importance, the true extent of their diversity has only recently been uncovered by molecular surveys. The diversity of picoeukaryotes has not yet been studied in the White Sea, which is a unique marine environment combining features of temperate and Arctic seas. Here, we investigated the taxonomic composition of eukaryotic picoalgae in ice and under-ice water at a station located in the Kandalaksha Bay of the White Sea. We applied metagenomic survey using Illumina sequencing. Eight main algae phyla, namely, Chlorophyta, Katablepharidophyta, Haptophyta, Dinophyta, Cercozoa, Bacillariophyta, Cryptophyta, and Ochrophyta were identified. The genera Paraphysomonas and Micromonas and the order Pedinellales were most numerous in plankton; the genera Paraphysomonas, Micromonas, and Metopion were most abundant in ice. The number of "rare" phylotypes was 80 in under-ice water and 112 in ice. Some taxa of nano- and microalgae are identified for the first time in the White Sea phytoplankton. Our data provide a basis for further research of tiny phototrophs in the Russian Arctic.

[RPS4 and NAD5 sequences evidenced of polyphyly of ditrichaceae and parallelisms in the evolution of haplolepidous mosses].

Molecular phylogenetic analysis of 66 representatives of haplolepidous mosses showed polyphylia of Ditrichaceae. According to the data obtained, the structure of the peristome, as well as features of the gametophyte on which a family traditionally allocated, arose independently in different groups of haplolepideous mosses. At least six genera (Distichium, Saelania, Eccremidium, Garckea, Rhamphidium, and Wilsoniella) should be excluded from the Ditrichaceae family, while Saelania and Distichium should be assigned even to another order. The loss of the peristome and forming of cleistocarpous capsules also occurs independently in at least two lineages of Ditrichaceae s. str., and in representatives of several lineages of Pottiaceae, a family derived from this group. Ditrichum, the type genus of Ditrichaceae, is also polyphyletic, species of this genus belong to two clades. It was concluded that parallel lines of the morphological variability in this group of mosses occur and its phylogeny need to be resolved based on molecular data.

TAS3 Genes for small ta-siARF RNAs in plants belonging to subtribe Senecioninae: occurrence of prematurely terminated RNA precursors.

The various classes of plant 21 - to 24-nt siRNAs derive from long dsRNA precursors that are processed by the ribonuclease Dicer-like (DCL). The species of ta-siRNA were originally discovered in Arabidopsis thaliana. Four gene families have been identified in Arabidopsis that each produces a number of ta-siRNAs: TAS1, TAS2, TAS3 and TAS4. The TAS3 genes encode tasiR-ARF species which target the mRNA of three Auxin Response Factor (ARF) genes (ARF2, ARF3/ETT and ARF4) for subsequent degradation. The function of TAS3 precursor RNA is controlled by two miR390 target sites flanking tandem of ta-siARF sequences. In this paper, we have studied the presence ofta-siARF RNA genes in the representatives of subtribe Senecioninae. Senecioneae is the largest tribe of Asteraceae, comprised of ca. 150 genera and 3,000 species which include many common succulents of greenhouses. Approximately one-third of species are placed in genus Senecio, making it one of the largest genera of flowering plants. However, there was no information on the structure of TAS genes in these plants. We revealed that the TAS3 species (TAS3-Sen1) in Senecio representatives was actively transcribed, and its homologues are distributed among many Asteracea plants and found to be similar to Arabidopsis AtTAS3a gene. We revealed several prematurely terminated transcripts of TAS3-Sen1. Finding the alternative shortened transcripts of TAS3-Sen1 lacking the 3'-terminal site cleaved by miR390 and retaining the 5'-terminal miR390 non-cleaved site suggested their using as decoys for the modulation of miR390 activity to regulate synthesis of ta-siARF RNAs in different Senecioninae species.

Novel miR390-dependent transacting siRNA precursors in plants revealed by a PCR-based experimental approach and database analysis.

TAS loci in plant genomes encode transacting small interfering RNAs (ta-siRNAs) that regulate expression of a number of genes. The function of TAS3 precursor in Arabidopsis thaliana is controlled by two miR390 target sites flanking two ta-siARF sequences targeting mRNAs of ARF transcription factors. Cleavage of the 3'-miR390-site initiates ta-siRNAs biogenesis. Here we describe the new method for identification of plant ta-siRNA precursors based on PCR with oligodeoxyribonucleotide primers mimicking miR390. The method was found to be efficient for dicotiledonous plants, cycads, and mosses. Based on sequences of amplified loci and a database analysis, a novel type of miR390-dependent TAS sequences was identified in dicots. These TAS loci are characterized by a smaller distance between miR390 sites compared to TAS3, a single copy of ta-siARF, and a sequence conservation pattern pointing to the possibility that processing of novel TAS-like locus is initiated by cleavage of the 5'-terminal miR390 target site.

Phagocytic activity of macrophages against liposomes with conjugates of oxidized dextrans and isonicotinic acid hydrazide during modeling of phagocytosis disturbances in vitro.

We studied phagocytic activity of macrophages against molecular-liposome hybrid compositions consisting of liposomes (diameter 200-450 nm) containing oxidized dextrans with a molecular weight of 35 or 60 kDa conjugated with the basic antituberculosis preparation isonicotinic acid hydrazide (dextrazides) during modeling of various disturbances of endocytosis function of phagocytic cells in vitro. Preincubation of macrophages with trypsin, colchicine, or sodium azide did not change the parameters of adhesion of molecular-liposome hybrid compositions to macrophages. It was found that preincubation of cells with colchicine or sodium azide reduced parameters of phagocytosis of the molecular-liposome hybrid compositions; this reduction did not depend on the molecular weight of dextrans entering the composition of the molecular-liposome hybrid compositions.

Effects of molecular liposomal hybrid compositions with oxidized dextrans and isonicotinic acid hydrazide on production of granulocytic macrophage colony-stimulating factor by macrophages.

The effects of molecular liposomal hybrid compositions consisting of liposomes (200-450 nm) containing oxidized dextrans (dextranals; 35-60 kDa) conjugated with isonicotinic acid hydrazide (dextrazides), their components, and native dextrans on the production of granulocytic macrophage CSF by peritoneal macrophages were studied in vitro. Dextranals proved to be more potent inductors of granulocytic macrophage CSF than native dextrans. Conjugation of nicotinic acid hydrazide with dextranals did not modify their capacity to stimulate the production of granulocytic macrophage CSF. Liposomes in the molecular liposomal hybrid compositions did not attenuate the dextrazide capacity to stimulate the production of granulocytic macrophage CSF. Molecular liposomal compositions containing 60 kDa dextrazide exhibited the most potent stimulatory effect on macrophage production of granulocytic macrophage CSF.

Phagocytosis of hybrid molecular nanosomal compositions containing oxidized dextrans conjugated with isonicotinic acid hydrazide by macrophages.

We studied phagocytic activity of macrophages towards hybrid molecular nanosomal compositions consisting of 150-800-nm nanoliposomes containing oxidized dextrans with a molecular weight of 35 and 60 kDa obtained by chemical ("permanganate") and radiochemical oxidation of dextran conjugated with isonicotinic acid hydrazide (dextrazides, intracellular prolonged antituberculous drugs). Phagocytic activity of macrophages towards hybrid molecular nanosomal compositions containing dextrazides obtained by chemical oxidation of dextrans is higher than activity towards hybrid molecular nanosomal compositions containing dextrazides prepared by radiochemical oxidation and depends on the size of hybrid molecular nanosomal compositions and molecular weight of oxidized dextrans.

In vitro effect of oxidized dextrans on peritoneal cells.

We studied the dependence of in vitro dextran biocompatibility on the method of oxidation of 35-kDa dextran. The biocompatibility of dextran oxidized with potassium permanganate was higher compared to that obtained by radiochemical oxidation. It was related to the formation of peroxide compounds during radiochemical oxidation.

Comparative study of the in vitro effect of nanoliposomes with oxidized dextrans on peritoneal cells.

We studied the in vitro effect of hybrid molecular-nanosomal biocompatible compositions on cultured peritoneal cells. The compositions consisted of oxidized dextrans with a mean molecular weight of 35 and 60 kDa, which were obtained by chemical and radiochemical oxidation of dextran. Hybrid nanoliposomal compositions of chemically oxidized dextran (permanganate method) had greater biocompatibility and tropic activity to macrophages compared to nanoliposomes of radiochemically oxidized dextran.

Comparative analysis of 5.8 S rRNA from Ephedra kokanica Regl. (Gymnospermae) and other plant species.

5.8 S rRNA from the gymnosperm Ephedra kokanica Regl. (EMBL Data Library accession No. X15676) has been sequenced. It is 161 nucleotides long and contains three 2'-O-methylated residues--two adenosines and one guanosine. No pseudouridine have been detected. E. kokanica 5.8 S rRNA, as those from other plant species, can form a secondary structure with paired 5'- and 3'-terminal regions. 5.8 S rRNAs of seed plants differ from the moss Mnium reguicum 5.8 S rRNA in that they have longer variable 'GC-rich' hairpins with insertions in the loop region. 5.8 S rRNA of E. kokanica reveals 69 and 82% of homology with that of moss and five angiosperm species, respectively. The posttranscriptional modification pattern of plant 5.8 S rRNAs is not strictly conservative.

Sequences of rDNA internal transcribed spacers from the chloroplast DNA of 26 bryophytes: properties and phylogenetic utility.

We determined the sequence of the region of the chloroplast DNA inverted repeat spanning from the 3'-terminus of the 23S rRNA gene to the 5'-terminus of the tRNA[Arg](ACG) gene (about 700 bp) from 25 bryophytes and from the charophycean alga Chara australis. Phylogenetic analysis of these sequences using the neighbor-joining method suggests an early dichotomy of bryophytes and their paraphyly relative to the tracheophyte lineage. A monophyly of liverworts (Marchantiidae plus Jungermanniidae), a deep divergence of Metzgeriales among Jungermanniidae and a close affinity of the two subclasses of mosses, Sphagnidae and Andreaeidae, are evident. The branching pattern observed is consistent with the phylogenetic distribution of several prominent indels observed in the alignment.

Experimental investigation of the mammary gland tumour phantom for multifrequency microwave radio-thermometers.

Microwave radiometry is a spectral measurement technique for resolving the electromagnetic radiation of matter when its temperature is above absolute zero. The radio-thermometer utilises this technique and consequently can provide temperature distributions in subcutaneous biological tissues. A new phantom was proposed that imitates a mammary gland tumour, and the brightness temperature was observed using radio-thermometers operated at different frequencies, 1.75GHz and 3GHz. The proposed phantom, simulating heat diffusion propagated by tissues around real tumours, revealed that the thermal characteristics of the tumour imitator were well matched to the heat transfer properties of a real tumour and a proportional linear relationship between the location of the tumour imitator and the brightness temperature in a homogenous medium was established. From experiments using the proposed mammary gland tumour phantom and a clinical trial on patients with breast cancer, it could be concluded that a radio-thermometer with a short wavelength (lambda = 10cm, i.e. f= 3GHz) is useful to resolve a thermal anomaly at a shallow depth in an homogenous medium such as a breast.