Gene expression analysis of primordial shoot explants collected from mature white spruce (Picea glauca) trees that differ in their responsiveness to somatic embryogenesis induction.

Within a plantation of clonal somatic embryo-derived white spruce trees that belonged to four genotypes, one genotype (G6) has consistently responded for the last 16 years, to the induction of somatic embryogenesis within primordial shoot explants. Analysis of fourteen individuals within this genotype subsequently revealed a group of clonal trees that were nonresponsive. This in turn provided a unique opportunity to conduct differential gene expression analysis in the absence of genotype-specific factors. Absolute qPCR was first used to expand the analysis of several genes previously identified via microarray analysis to be differentially expressed during SE induction, along with the inclusion of two nonresponsive genotypes. While this demonstrated a high level of repeatability within, and between, responsive and nonresponsive genotypes, it did not support our previous contention that an adaptive stress response plays a role in SE induction responsiveness, at least with respect to the candidate genes we analyzed. RNAseq analysis was then used to compare responsive and nonresponsive G6 primordial shoots during the somatic embryogenesis induction treatment. Although not analyzed in this study, this included samples of callus and embryonal masses previously generated from G6 explants. In addition to revealing a large number of differentially expressed genes, de novo assembly of unmapped reads was used to generate over 25,000 contigs that potentially represent previously unidentified transcripts. This included a MADS-domain gene that was found to be the most highly differentially expressed gene within responsive shoot explants during the first seven days of the induction treatment.

Organ Donation: What Are the Opinions of Nursing Students at the University of Bialystok in Poland?

BACKGROUND: Nursing students, as future professionals, have an enormous impact on the opinion of the rest of the population on topics of organ donation and transplantation (ODT). The objective of this study was to analyze the attitudes of nursing students at the University of Bialystok toward ODT, and to determine the factors that affect ithemt. METHODS: Academic year 2011-2012. STUDY POPULATION: nursing students at the University of Bialystok. Type of sampling: sampling points during 5 compulsory-attendance nursing courses with >80% fulfillment. Measuring instrument: validated questionnaire (PCID-ODT Rios) filled out anonymously and self-administered. RESULTS: Completion rate of 90% (331/367): 58% (n = 191) in favor, 35% (n = 118) undecided, and 7% (n = 22) against. The most important variables were: being in the 1st year (odds ratio [OR], 4.444; 95% confidence interval [CI], 11.627-1.700; P = .002); not knowing any donor (OR, 12.048; 95% CI, 55.555-2.57; P = .018); being in favor of organ donation by a relative (OR, 6.25; 95% CI, 14.705-2.666; P < .001); discussing the topic with friends (OR, 2,785; 95% CI, 5.347-1.447; P = .002); not worried about the possible mutilation of the body after donation (OR, 3.039; 95% CI, 5.882-1.569; P = .001); favorable attitude toward ODT of mother (OR, 3.676 (0.881-1.814; P < .001); accepting other treatment options, such as inhumation (OR, 3.436; 95% CI, 7.092-1.655; P = .001); accepting autopsy after death (OR, 2.141; 95% CI, 4.065-1.126; P = .020); and professing the Catholic religion (OR, 3.436; 95% CI, 8.333-1.410; P = .007). CONCLUSIONS: More than 42% of the nursing students were not in favor of organ donation, their attitudes influenced by various psychosocial factors.

Gene expression profiling of shoot-derived calli from adult radiata pine and zygotic embryo-derived embryonal masses.

BACKGROUND: Although somatic embryogenesis has an unprecedented potential for large-scale clonal propagation of conifers, the ability to efficiently induce the embryonal cultures required for somatic embryo production has long been a challenge. Furthermore, because early stage zygotic embryos remain the only responsive explants for pines, it is not possible to clone individual trees from vegetative explants at a commercial scale. This is of particular interest for adult trees because many elite characteristics only become apparent following sexual maturation. FINDINGS: Shoot explants collected from adult radiata pine trees were cultured on four induction media differing in plant growth regulator composition, either directly after collection or from in vitro-generated axillary shoots. Six callus lines were selected for microscopic examination, which failed to reveal any embryonal masses (EM). qPCR expression profiling of five of these lines indicated that explant type influenced the absolute level of gene expression, but not the type of genes that were expressed. The analysis, which also included three EM lines induced from immature zygotic embryos, encompassed five categories of genes reflective of metabolic, mitotic and meristematic activity, along with putative markers of embryogenicity. Culture medium was found to have no significant impact on gene expression, although differences specific to the explant's origin were apparent. Expression of transcriptional factors associated with vegetative meristems further suggested that all of the callus lines possessed a substantive vegetative character. Most notable, however, was that they all also expressed a putative embryogenic marker (LEC1). CONCLUSIONS: While limited in scope, these results illustrate the utility of expression profiling for characterizing tissues in culture. For example, although the biological significance of LEC1 expression is unclear, it does present the possibility that these callus lines possess some level of embryogenic character. Additionally, expression of vegetative meristem markers is consistent with their vegetative origin, as are differences in expression patterns as compared with EM.

Seasonal variation in ischaemic stroke frequency in Podlaskie Province by season.

PURPOSE: The aim of the present study was to assess seasonal differences in ischaemic stroke among patients hospitalized in Department of Neurology in Bialystok during 2002-2005. MATERIAL AND METHODS: To examine the seasonal incidence of ischaemic stroke, we analyzed data from the Department of Neurology in Bialystok in a retrospective study. The year was divided into four seasons: spring (March, April, May), summer (June, July, August), autumn (September, October, November), and winter (December, January, February). Seasonal differences were studied in relation to the following clinical characteristics: age, gender, history of stroke, and time of stroke onset. RESULTS: Age of patients with ischaemic stroke ranged 19 between 101 years, a mean age was 72.4 +/- 12 years. Incidence of ischaemic stroke increased in the last years. We noted a higher incidence of ischaemic stroke in older patients (mean 74.36 years old) during winter months than in patients (71.40 years) in summer months. Gender had no effect on incidence of ischaemic stroke. Significant seasonal variation of ischaemic stroke in all years (p = 0.0010) and for 2005 year (0.0090) were found. Incidence of ischaemic stroke was depend on month of year. Significant increase of ischaemic stroke was noted in December. The lowest incidence of stroke was observed in August and September. CONCLUSION: Incidence of ischaemic stroke increased in the last years. The present findings suggest an increase in the incidence of ischaemic stroke in winter in December.

Comparative analysis of quality of life women in menopause period in Poland, Greece and Belorussia using MRS Scale. Preliminary report.

PURPOSE: The aim of this study was the assessment of climacteric symptoms, the activity and quality of life of women in menopausal period from Poland, Greece and Belorussia using a Menopause Rating Scale (MRS). MATERIAL AND METHODS: The study was conducted among women in age after 45 years, from Poland (55), Belorussia (50) and Greece (85). MRS was obtained from the Professor Heinemann from Center of Epidemiology and Health Studies in Berlin. The scoring scheme is simple, i.e. the score increases point by point with increasing severity of subjectively perceived symptoms in each of the 11 items (severity 0--no complaints, 4 scoring points--severe). The respondent provides her personal perception by checking one of 5 possible boxes of "severity" for each of the items. RESULTS: Mild and no complaints in similar degree were reported by all women from these three countries. We found significant (p < 0.001) differences between severe complaints reported by Greek women compared with complaints respondents from Belorussia and Poland. Moderate complaints were reported more frequently by women from Poland (32.56%) and Belorussia (34%) compared with women from Greece (28.55%). Severe complaints were noted more rarely in 1.6% Greek women compared with 2.6% Belorussian and 3% Polish respondents. No significant differences between no complainants, mild, moderate, marked and severe between women from Belorussia, Poland and Greece. CONCLUSIONS: Generally we did not observe significant differences between reported complaints by women from Belorussia, Poland and Greece.

Exposure the doctors to aggression in the workplace.

PURPOSE: To evaluate the frequency, source and type of aggression towards doctors, depending on their place of work and position. MATERIAL AND METHODS: The study was conducted among 501 doctors from the area of Podlaskie Province. To evaluate the level and type of aggression towards doctors in their workplace we used a questionnaire prepared for the needs of this study by modifying the questionnaire "The frequency and consequences of exposing nurses to workplace aggression", which had been drafted by the Institute of Labour Medicine in Lódz. The results were analysed with the application of the chi-square and the Kruskal-Wallis tests. RESULTS: The most common form of aggression was voice raising, which happened to 80% of doctors employed in inpatient medical centres and 91% doctors from outpatient centres. More than a half of the subjects have heard threats from their patients. Verbal aggression from doctors' superiors happened most often in surgery wards (48%), neurology wards (40%), admission rooms (33%). The causes of aggression most often quoted by doctors include: staff shortages (9%), stress--tiredness (9%). CONCLUSIONS: Workplace aggression towards doctors may be inflicted both by patients and colleagues. The aggression in the medical environment can take on different forms and create a threat in the workplace. Doctors working in hospital wards (psychiatry, surgery, neurology) are the ones who are the most exposed to aggression.

Transgene integration patterns and expression levels in transgenic tissue lines of Picea mariana, P glauca and P abies.

Seventy transgenic tissue lines (translines) of three spruce species ( Picea mariana, P. glauca and P. abies) were characterized with respect to the integration pattern of the gus (beta-glucuronidase) gene, and the level of GUS activity was determined in 81 lines. The majority of the P. mariana translines (18 lines of 22) integrated multicopies of the transgene, whereas mostly single integrations were detected in the other two species. The activity levels of GUS varied widely among the individual translines of P. mariana, and there was a strong indication that the logarithm of GUS activity increased with the number of gus copies ( P=0.0003) in lines with one to five known insertions (uncensored). The average level of GUS activity, in lines that integrated one gene copy, was the highest in white spruce followed by black spruce and Norway spruce (22.7, 16.5 and 6.3 nmol 4-methylumbelliferone min(-1 )mg(-1 )protein, respectively).

[The sorption of polycyclic aromatic hydrocarbons in Zegrzynskie Lake]. / Sorpcja wielopierscieniowych weglowodorów aromatycznych w osadach dennych pochodzacych z Jeziora Zegrzynskiego.

The sorption of polycyclic aromatic hydrocarbons in aquatic sediments from Zegrzynskie Lake was examined. Batch experiment was performed in order to determine sorption efficiency in different kinds of sediments from Zegrzynskie Lake. Five polycyclic aromatic hydrocarbons (phenanthrene, fluoranthene, pyrene, chrysene benzo[a]pyrene) were chosen to this experiment and sorption process was examined on seven sediments of different properties. Chosen hydrocarbons are of different structure of molecule and different chemical and physical properties. Concentrations of polycyclic aromatic hydrocarbons in aquatic sediments and in water phase were measured in the following order: extraction with dichloromethane, concentration on rotary evaporator, silica gel clean up, n-hexane elution, concentration on rotary evaporator and in vials, GC/MS analysis. Chemical composition of aquatic sediments were examined using methods for sewage sludge and soils analysis. In every sediment concentrations of PAHs, organic matter and organic: carbon, phosphorus, nitrogen and sulphur were measured. Also fractional analysis of sediments was made. Isotherms of sorption were measured for these sediments and compounds. Equations of these isotherms were performed and were used in order to find relationships between sorption efficiency and sediments composition. Depending on sediment properties and composition different concentrations of polycyclic aromatic hydrocarbons were found in solid phase. Sediments of high quantities of organic matter and small particles were the best sorbents for PAHs. Fluoranthene, pyrene and chrysene were efficiently sorbed in sediments of high concentration of organic matter. And efficiency of phenanhrene and benzo[a]pyrene sorption were better in sediments with high quantity of organic sulphur.

Cytological and molecular relationships between Larix decidua, L. leptolepis and Larix x eurolepis: identification of species-specific Chromosoms and synchronization of mitotic cells.

The effects of different concentrations of hydroxyurea (HU) and aphidicolin (APH) on the mitotic index (MI) were compared in cells of embryogenic cultures of Larix decidua, L. leptolepis, and L. decidua x L. leptolepis (Larix x eurolepis). The highest enhancement of the MI was obtained with HU at 1.25 mM and 0.6% colchicine. In general the MI decreased with an increase of HU or APH concentration (over 1.25 mM for HU and 5 µM for APH). Detailed karyotype analyses were made on the somatic complement of L. decidua, L. leptolepis, and their hybrid. These karyotypes were asymmetrical and advanced, with the smaller chromosomes being more submedian than the larger ones. The topography of chromosome 7 of L. decidua and chromosome 9 of L. leptolepis was found to be the most significant cytotaxonomic characteristic in differentiating these two species. Cytological data indicate that Japanese larch (L. leptolepis) is phylogenetically closer to European larch (L. decidua) than the Siberian larch group (L. sibirica and L. sukaczewii). Chromosomes with unusually long kinetochores were found in both species and the hybrid. Hyperploid cells (2n = 25) were observed in the hybrid (Larix x eurolepis) material analyzed. A genomic L. decidua probe hybridized strongly to dots of DNA from L. leptolepis indicating that there is high sequence homology between these two species.

Genetic variation in somatic embryogenic response in open-pollinated families of black spruce.

Zygotic embryos from open-pollinated seeds of 20 black spruce (Picea mariana) families were used to investigate the proportion of genotypes that would give rise to embryogenic tissue (ET) and mature somatic embryos. Eighty-five percent of the maternal genotypes gave rise to embryogenic tissue. Within-family rates of ET induction ranged from 0 to 17%, with an average of 8%. The largest proportion of variation was among families, indicating the additive nature of the genetic variation. On a medium with 6% sucrose and 3.7 µM ABA, 90% of the embryogenic lines gave rise to abundant (>100/100 mg of ET), well-formed, mature somatic embryos. A medium with 2% sucrose, without 2,4-D, was used to germinate the mature somatic embryos. These were grown in the greenhouse and have now been established in field trials.

Recovery of somatic embryos and plantlets from protoplast cultures of Larix × eurolepis.

Somatic embryos and plantlets were regenerated from protoplasts of hybrid larch (Larix × eurolepis) isolated from two embryogenic callus and cell suspension culture lines (L1 and L2). L2, which was highly embryogenic, consistently yielded protoplasts that gave rise to somatic embryos. Centrifugation on a discontinuous medium/Percoll density gradient resulted in accumulation of embryogenic protoplasts in one of the Percoll interfaces. First division frequencies were in the range of 28-39% in line 1 and 18-20% in line 2 in both liquid and agarose-solidified culture media. The critical factor in maintaining high viability of cultures was lowering of osmotic pressure by dilution of the initial medium. The first somatic embryos were detected in 23- to 28-day-old cultures. Some of these developed into plants that were transferred to soil.