Ovarian transcriptome analysis of diploid and triploid rainbow trout revealed new pathways related to gonadal development and fertility.

Triploidisation represents several advantages (e.g. sterility) and therefore is routinely applied in aquaculture of several commercially important fish species, including rainbow trout. The comparative transcriptomic analysis of ovaries of triploid (3N) and diploid (2N) female rainbow trout revealed a total of 9 075 differentially expressed genes (DEGs; 4 105 genes upregulated in 2N and 4 970 genes upregulated in 3N ovaries, respectively). Identified clusters for DEGs upregulated in 3N and 2N ovaries were different, including carbohydrate and lipid metabolic process and transport, protein modification, signalling (related to folliculogenesis) and response to stimulus for DEGs upregulated in 2N, and developmental process, signalling (related to apoptosis, cellular senescence and adherence junctions) and regulation of RNA metabolic process for DEGs upregulated in 3N. The enrichment of processes involved in carbohydrate and lipid metabolism in 2N ovaries indicated high metabolism of ovarian tissue and the energy reservoir generation indispensable during the earliest stages of development. Our results highlight the importance of oocyte hydration along with oestrogen, insulin, leptin, fibroblast growth factor, and Notch signalling and pathways related to the regulation of cyclic adenosine monophosphate (cAMP) levels in proper oocyte meiotic maturation prior to ovulation in 2N ovaries. Conversely, triploidisation may lead to an increase in ovarian cellular senescence and apoptosis, which in turn can result in abnormal gonadal morphology and fibrosis. The downregulation of genes responsible for the precise regulation of meiosis and proper chromosome segregation during meiosis probably affects meiotic maturation via irregular meiotic division of chromosomes. The induction of triploidy of the rainbow trout genome resulted in enhanced expression of male-specific genes, genes responsible for re-establishing the transcriptional balance after genome reorganisation and genes involved in regulatory mechanisms, including gene silencing and DNA methylation. To the best of our knowledge, this is the first genome-wide investigation providing in-depth comprehensive and comparative gene expression patterns in the ovary from 2N and 3N rainbow trout females helping in elucidating the molecular mechanisms leading to impaired gonadal development and sterility of female triploids.

Exposure to the electromagnetic field alters the transcriptomic profile in the porcine endometrium during the peri-implantation period.

A low-frequency electromagnetic field (EMF) is an environmental pollutant that may influence female reproduction. This research was undertaken to test the hypothesis that EMF causes alterations in the transcriptomic profile of the endometrium. This study investigated the in vitro effects of EMF treatment (50 Hz, 2 h) on global transcriptome alterations in the endometrium isolated from pigs during the peri-implantation period. The control endometrium was not treated with EMF. The EMF treatment altered the expression of 1561 transcriptionally active regions (TARs) in the endometrium. In the group of 461 evaluated DEGs, 156 were up-regulated (34%), 305 were down-regulated (66%) and 341 (74%) had known biological functions. A total of 210 long noncoding RNAs (lncRNAs) with changes in expression profiles, and 146 predicted RNA editing sites were also evaluated. Exposure to EMF changes the expression of genes encoding proteins that are involved in proliferation and metabolism in endometrial tissue. These results provide useful inputs for further research into the impact of EMF on molecular changes in the uterus during the peri-implantation period and, consequently, pregnancy outcome.

Transcriptome analysis of boar spermatozoa with different freezability using RNA-Seq.

Semen freezability is associated with genetic markers, and there is a diverse set of sperm transcripts that have been attributed to various cellular functions. RNA-Seq was performed to compare the transcript profiles of spermatozoa from boars with different semen freezability. We examined ejaculates from the Polish large white (PLW) boars that were classified as having good and poor semen freezability (GSF and PSF, respectively; n = 3 boars per group) by assessing post-thaw motility characteristics, mitochondrial membrane potential, plasma membrane and acrosome integrity. Total RNA was isolated from fresh spermatozoa from boars of the GSF and PSF groups and subjected to RNA-Seq (Illumina NextSeq 500 platform). Transcript abundance was assessed with the DESeq2, DESeq, and EdgeR Bioconductor R packages, and varying numbers of differentially expressed gene (DEG) transcripts were detected in the spermatozoa of each boar. Using RNA-Seq, we identified several genes associated with inflammation and apoptosis (FOS, NFATC3, ITGAL, EAF2 and ZDHHC14), spermatogenesis (FGF-14 and BAMBI), autophagy (RAB33B), protein phosphorylation (PTPRU and PTPN2) and energy metabolism (ND6 and ACADM) that were predominantly up-regulated in poor freezability ejaculates. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) validated the transcript expression levels detected by RNA-Seq and thus confirmed the reliability of this technique. Subsequent validation with western blotting showed that the expression of three proteins was in accordance with the transcript abundance. Overall, we demonstrated that the up-regulation of the DEG transcripts in spermatozoa was associated with poor semen freezability. We suggest that spermatozoa transcriptome profiling provides a foundation to further elucidate the relevance of sperm-related transcripts on cryo-survival. The sperm-related transcripts, namely FOS, NFATC3, EAF2, BAMBI, PTPRU, PTPN2, ND6 and ACADM, are potential markers for predicting the freezability of boar semen.

Identification of differentially expressed placental transcripts during multiple gestations in the Eurasian beaver (Castor fiber L.).

The Eurasian beaver is one of the largest rodents that, despite its high impact on the environment, is a non-model species that lacks a reference genome. Characterising genes critical for pregnancy outcome can serve as a basis for identifying mechanisms underlying effective reproduction, which is required for the success of endangered species conservation programs. In the present study, high-throughput RNA sequencing (RNA-seq) was used to analyse global changes in the Castor fiber subplacenta transcriptome during multiple pregnancy. De novo reconstruction of the C. fiber subplacenta transcriptome was used to identify genes that were differentially expressed in placentas (n=5) from two females (in advanced twin and triple pregnancy). Analyses of the expression values revealed 124 contigs with significantly different expression; of these, 55 genes were identified using MegaBLAST. Within this group of differentially expressed genes (DEGs), 18 were upregulated and 37 were downregulated in twins. Most DEGs were associated with the following gene ontology terms: cellular process, single organism process, response to stimulus, metabolic process and biological regulation. Some genes were also assigned to the developmental process, the reproductive process or reproduction. Among this group, four genes (namely keratin 19 (Krt19) and wingless-type MMTV integration site family - member 2 (Wnt2), which were downregulated in twins, and Nik-related kinase (Nrk) and gap junction protein ß2 (Gjb2), which were upregulated in twins) were assigned to placental development and nine (Krt19, Wnt2 and integrin α7 (Itga7), downregulated in twins, and Nrk, gap junction protein ß6 (Gjb6), GATA binding protein 6 (Gata6), apolipoprotein A-I (ApoA1), apolipoprotein B (ApoB) and haemoglobin subunit α1 (HbA1), upregulated in twins) were assigned to embryo development. The results of the present study indicate that the number of fetuses affects the expression profile in the C. fiber subplacental transcriptome. Enhancement of transcriptomic resources for C. fiber will improve understanding of the pathways relevant to proper placental development and successful reproduction.

The complete mitogenome of Colletotrichum lupini var. setosum.

The structure of Colletotrichum lupini mitogenome is typical of a fungus from the genus Colletotrichum similar to C. acutatum and C. lindemuthianum. The sequenced mitogenome has a total length of 36 554 bp. The nucleotide composition in the following genome is: 35.7% - A, 16.5% - C, 13.4% - G and 29.9% - T. In the C. lupini mitogenome we identified 46 genes: 15 protein coding genes, two ribosomal RNAs and 29 tRNA genes.

The opening ceremony of the Radiopharmaceuticals Production and Research Centre at the Heavy Ion Laboratory of the University of Warsaw, May 15, 2012. Followed by an International Conference PETRAD2012.

Two important events happened at the Heavy Ion Laboratory of the University of Warsaw in May this year. First, on May 15 the new Radiopharmaceuticals Production and Research Centre (RPRC) was inaugurated by the Rector Elect, Prof. Marcin Palys. The construction of this Centre, located on the premises of the Laboratory, was supported by grants from the Ministry of Sciences, International Atomic Energy Agency, Ministry of Health and European Structural Funds.

Interferon-gamma release assays are a better tuberculosis screening test for hemodialysis patients: A study and review of the literature.

Diagnosing latent tuberculosis (TB) infection (LTBI) in dialysis patients is complicated by poor response to tuberculin skin testing (TST), but the role of interferon-gamma release assays (IGRAs) in the dialysis population remains uncertain. Seventy-nine patients were recruited to compare conventional diagnosis (CD) with the results of two IGRA tests in a dialysis unit. Combining TST, chest x-ray and screening questionnaire results (ie, CD) identified 24 patients as possible LTBI. IGRA testing identified 22 (QuantiFERON Gold IT, Cellestis, USA) and 23 (T-spot.TB, Oxford Immunotec, United Kingdom) LTBI patients. IGRA and CD correlated moderately (κ=0.59). IGRA results correlated with history of TB, TB contact and birth in an endemic country. TST was not helpful in identifying LTBI patients in this population. The tendency for IGRAs to correlate with risk factors for TB, active TB infection and history of TB argues for their superiority over TST in dialysis patients. There was no superiority of one IGRA test over another.

Le diagnostic d'infection tuberculeuse latente (ITBL) chez les patients sous dialyse est compliqué par le peu de réponse au test cutané à la tuberculine (TCT), mais le rôle du test de libération d'interféron gamma (TLIG) au sein de la population sous dialyse demeure incertain. Les auteurs ont recruté 79 patients pour comparer le diagnostic classique (DC) aux résultats de deux TLIG au sein d'une unité de dialyse. L'association du TCT, de la radiographie pulmonaire et des résultats d'un questionnaire de dépistage (c.-à-d. le DC) a permis de dépister 24 patients comme pouvant être atteints d'une ITBL. Le TLIG a permis de dépister 22 (QuantiFERON Gold IT, Cellestis, États-Unis) et 23 (T-spot.TB, Oxford Immunotec, Royaume-Uni) patients atteints d'une ITBL. Le TLIG et le DC avaient une corrélation modérée (κ=0,59). Les résultats du TLIG étaient corrélés avec les antécédents de tuberculose (TB), les contacts atteints de TB et la naissance dans un pays endémique. Le TCT ne contribuait pas à dépister les patients atteints d'une ITBL au sein de cette population. La tendance des TLIG à être corrélés avec les facteurs de risque de TB, une infection active par la TB et les antécédents de TB laisse supposer leur supériorité par rapport au TCT chez les patients sous dialyse. Aucun type de TLIG n'était supérieur aux autres.

Parenteral vitamin B12 in macrocytic hemodialysis patients reduced MMA levels but did not change mean red cell volume or hemoglobin.

AIMS: Unexplained macrocytic anemia was common in our hemodialysis (HD) unit. Vitamin B12 requirements may be higher in HD patients; therefore, patients may be deficient despite "normal" serum levels. We studied vitamin B12 status and the effect of parenteral vitamin B12 administration in macrocytic HD patients. A normocytic group was included for comparison. MATERIALS AND METHODS: Prospective cohort study of 62 HD patients (34 macrocytic, 28 normocytic) from November 2008 to March 2009. Patients were on stable doses of darbepoetin and iron replete. Vitamin B12 1,000 µg IV was given once weekly for 4 weeks and follow-up was 12 weeks. Methylmalonic acid (MMA) level was used as an indicator of vitamin B12 status. MCV and hemoglobin were also examined for an effect of B12 administration. RESULTS: At baseline: all patients had serum B12 levels > 200 pmol/l; 97% had serum folate levels > 55 nmol/l; there was no difference in serum B12 levels between groups (504 vs. 571 pmol/l, p = 0.18); MMA was higher in the macrocytic group (0.56 vs. 0.48 µmol/l, p = 0.048) and hemoglobin (Hg) was lower (119 vs. 125 g/l, p = 0.03); median darbepoetin dose was equivalent (20 µg/week). Following IV vitamin B12, the macrocytic group had a greater and more sustained reduction in MMA (-0.064 vs. -0.0066 µmol/l/wk, p = 0.004). There was no improvement in hemoglobin (Hg), reticulocyte count or MCV in either group. Median darbepoetin dose was unchanged. CONCLUSIONS: IV vitamin B12 led to a sustained decline in MMA levels in macrocytic patients, suggesting functional vitamin B12 deficiency at baseline. However, there were no significant changes in Hg or darbepoetin dose.

Complete switch to darbepoetin in a hemodialysis unit.

AIMS: In March 2003, our hemodialysis unit switched all patients from subcutaneous (s.c.) rHuEPO to intravenous (i.v.) darbepoetin. The primary outcome was to assess the efficacy of i.v. darbepoetin to maintain target serum hemoglobin (Hb) compared to s.c. rHuEPO. Secondary outcomes were to evaluate the manufacturer's recommend guidelines for conversion of rHuEPO to darbepoetin, and to assess the cost implications of darbepoetin therapy. METHODS: This was an 18-month open-label observational study of 95 hemodialysis patients. At the time of the switch to darbepoetin (baseline), data were collected retrospectively for six months and prospectively for 12 months, at three-month intervals. The first six months of darbepoetin therapy was considered a dose titration phase, thus, data were analyzed comparing two six-month periods: (-) six months to baseline (rHuEPO phase) and (+) 6-12 months (darbepoetin phase). Doses were titrated to a target Hb of 120-135 g/l. RESULTS: There was no significant difference in Hb between phases at any time point. Mean Hb ranged from 119.6-121.5 g/l for rHuEPO and 121.9-123.4 g/l for darbepoetin. The median darbepoetin dose remained stable throughout the analysis at 30 microg/week while the median dose of rHuEPO rose from 8,000 U/week at minus six months to 9,000 U/week at baseline. Median 12-month cost savings associated with the administration of darbepoetin were estimated at 212,000 dollars. The recommended darbepoetin dose from the manufacturer's conversion table was deemed too low for baseline rHuEPO doses above 17,000 U/week. A more simplified dose conversion nomogram was created. CONCLUSION: Darbepoetin was able to maintain similar serum Hb levels compared to rHuEPO at a substantially reduced cost.

Efficacy of low-dose alteplase for treatment of hemodialysis catheter occlusions.

BACKGROUND: Traditionally, alteplase 2 mg/lumen doses have been used to treat central venous catheter (CVC) occlusions. On January 20, 2004, our hemodialysis (HD) unit implemented a new protocol to utilize alteplase 1 mg/lumen doses for catheter occlusion. OBJECTIVES: The objectives were to 1) assess the efficacy of low-dose alteplase in restoring HD catheter patency; 2) determine the duration of CVC patency as determined by need for further alteplase doses, or radiological or surgical line interventions; and 3) evaluate the financial implications of the new protocol. METHODS: The study was a prospective, open-label trial of 50 consecutive HD patients with permanent, tunnelled CVC lines. A treatment course consisted of 1 or 2 doses of alteplase instilled for 60 minutes then aspirated or as an overnight (48-72 hour) dwell until the next HD. The patient's first alteplase dose following implementation of the new protocol was evaluated. Patients were followed for two months to record need for further al-teplase treatment courses, and four months to document radiological or surgical line interventions. The primary outcome was to assess successful restoration of catheter patency defined as the ability of alteplase to restore or maintain HD blood flow rate at or above 300 mL/minute. A financial analysis compared alteplase costs for 11 months prior to and after implementation of the new protocol. RESULTS: Alteplase 1 mg/lumen doses restored catheter patency in 72% of HD patients with one dose, increasing to 83% with a second dose. Sixty-two percent of patients required a subsequent alteplase course with a median time to next treatment of 14 days and a median of 2 courses/patient. Radiological interventions were ordered in 38% of patients resulting in 8 lines replacements and 7 line strippings. Financial savings with the new low-dose protocol were approximately CDN dollar 22,000. CONCLUSION: Low dose alteplase 1 mg/lumen successfully treated occlusion of permanent hemodialysis catheters, with a resulting cost reduction.

Neutron density distributions deduced from antiprotonic atoms.

The differences between neutron and proton density distributions at large nuclear radii in stable nuclei were determined. Two experimental methods were applied: nuclear spectroscopy analysis of the antiproton annihilation residues one mass unit lighter than the target mass and the measurements of strong-interaction effects on antiprotonic x rays. Assuming the validity of two-parameter Fermi neutron and proton distributions at these large radii, the conclusions are that the two experiments are consistent with each other and that for neutron rich nuclei it is mostly the neutron diffuseness which increases and not the half-density radius. The obtained neutron and proton rms radii differences are in agreement with previous results.

[A case of bilateral, simultaneous breast amputation for cancer palpated in the left breast and undiagnosed in the right breast with different histologic classifications]. / Przypadek obustronnej, jednoczasowej amputacji gruczolu piersiowego z powodu badalnego palpacyjnie raka piersi lewej i niebadalnego piersi prawej o róznej klasyfikacji histologicznej.

The paper presents the case of bilateral simultaneous breast amputation due to different histological classification cancer. The right breast tumor was non-palpable.

Functionalized carbosilane dendritic species as soluble supports in organic synthesis.

A new methodology, which is compatible with the use of reactive organometallic reagents, has been developed for the use of carbosilane dendrimers as soluble supports in organic synthesis. Hydroxy-functionalized dendritic carbosilanes Si[CH2CH2CH2SiMe2(C6H4CH(R)OH)]4 (G0-OH, R = H or (S)-Me) and Si[CH2CH2CH2Si[CH2CH2CH2SiMe2(C6H4CH(R)OH)]3]4 (G1-OH, R = H or (S)-Me) were prepared and subsequently converted into the esters Si[CH2CH2CH2SiMe2(C6H4CH(R)OC(O)CH2Ph)]4 (R = H or (S)-Me) and Si[CH2CH2CH2Si[CH2CH2CH2SiMe2(C6H4CH(R)OC(O)CH2C6H4 R')]3]4 (R = H and R' = H or R = (S)-Me and R' = H or R = H and R' = Br). As an example the latter compound was functionalized under Suzuki conditions. The functionalized carboxylic acid was obtained in high yield after cleavage from the dendritic support. Moreover, the ester functionalized dendrimers were converted to the corresponding zinc enolates followed by a condensation reaction with an imine to a beta-lactam in excellent yield and purity. Furthermore, it was demonstrated that a small combinatorial library of beta-lactams could be prepared starting from a carbosilane dendrimer functionalized with different ester moieties. These results show that carbosilane dendrimers can be applied as soluble substrate carriers for the generation of low molecular weight organic molecules. In combination with nanofiltration techniques, separation and recycling of the dendrimers can be realized.

Evaluation of the effect of intravenous l-carnitine on quality of life in chronic hemodialysis patients.

AIM: The aim of this study was to determine the effect of l-carnitine on quality of life (QOL) in chronic hemodialysis patients. PATIENTS AND METHODS: This trial used a randomized, prospective, placebo-controlled, double-blind, crossover design. Inclusion criteria were patients who were older than 18 years, had been on dialysis for a minimum of one year, and had at least two of the following symptoms: intradialytic hypotension, muscle cramping, lack of energy, muscle weakness or myopathy, cardiomyopathy, or lack of responsiveness to erythropoietin (EPO). Patients were excluded if they were mentally incompetent to complete a QOL questionnaire. Sixteen patients were randomized to receive either l-carnitine (20 mg/kg) or placebo (normal saline) after each dialysis session for 12 weeks, followed by a 6-week washout, then the crossover therapy for 12 weeks. The Kidney Dialysis Questionnaire was the assessment tool used to evaluate QOL. RESULTS: There was no significant effect of l-carnitine on QOL irrespective of treatment order. There were also no differences found in any of the secondary outcomes including incidence of muscle cramping, intradialytic hypotension, EPO requirements or hemoglobin. Adverse effects consisted of gastrointestinal symptoms, with a similar incidence between l-carnitine and placebo. CONCLUSION: L-carnitine did not have a benefit on QOL in our patient population.