High resolution physical mapping of single gene fragments on pachytene chromosome 4 and 7 of Rosa

Background: Rosaceae is a family containing many economically important fruit and ornamental species. Although fluorescence in situ hybridization (FISH)-based physical mapping of plant genomes is a valuable tool for map-based cloning, comparative genomics and evolutionary studies, no studies using high resolution physical mapping have been performed in this family. Previously we proved that physical mapping of single-copy genes as small as 1.1 kb is possible on mitotic metaphase chromosomes of Rosa wichurana using Tyramide-FISH. In this study we aimed to further improve the physical map of Rosa wichurana by applying high resolution FISH to pachytene chromosomes. Results: Using high resolution Tyramide-FISH and multicolor Tyramide-FISH, 7 genes (1.7-3 kb) were successfully mapped on pachytene chromosomes 4 and 7 of Rosa wichurana. Additionally, by using multicolor Tyramide-FISH three closely located genes were simultaneously visualized on chromosome 7. A detailed map of heterochromatine/euchromatine patterns of chromosome 4 and 7 was developed with indication of the physical position of these 7 genes. Comparison of the gene order between Rosa wichurana and Fragaria vesca revealed a poor collinearity for chromosome 7, but a perfect collinearity for chromosome 4. Conclusions: High resolution physical mapping of short probes on pachytene chromosomes of Rosa wichurana was successfully performed for the first time. Application of Tyramide-FISH on pachytene chromosomes allowed the mapping resolution to be increased up to 20 times compared to mitotic metaphase chromosomes. High resolution Tyramide-FISH and multicolor Tyramide-FISH might become useful tools for further physical mapping of single-copy genes and for the integration of physical and genetic maps of Rosa wichurana and other members of the Rosaceae

Search for excess showers from Crab Nebula

The arrival directions of muon poor showers registrated in the Tien Shan experiment during an effective running time about I,8.IO(4)h were analyzed. It is shown that there is a significant excess of these showers coming the direction of Crab Nebula

Ecological Assessment of differrent raw materials for biogas

ABSTRACT: A comparative ecologic valuation of different raw materials for biogas production has been made. The valuation is based on the qualities of the different substrates: from energy cultures (sugar, fodder and red beet, leaves mass from the Paulovnia spring and autumn leaves). Three type of fertilizer (cattle from non-litter breeding, swine and non-removable bird fertilizer litter) as well as substrates from waste water of the WTP- Waster water treatment plan (mixed sediment, primary and secondary sediment). The examined raw material has been analyzed according to the following parameters: Dry matter (DM), Organic matter (OM), Organic Carbon (OC), Nitrogen Kjeldahl (N), Nitrogen Ammonium (N - ammonium), Nitrogen nitrate ( N- nitrate), Patassium (K), P (Phosphorus), pH ( H2O) and heavy metals – arsenic (As), Cadmium (Cd), Chromium (Cr), Copper (Cu), Mercury (Hg), Nickel (Ni), Lead (Pd), Zinc (Zn). The results of the examined substrates received from different raw materials (vegetal, organic fertilizers and waste water) give us ground to make the following conclusions: the examined substrates respond in the physical-chemical parameters to all technological requirements of raw material for biogas production (DM,C and proportion C:N). The established differences in the content of nitrogen / within the borders of 1,8% (substrate-leaves mass Paulovnia- spring) to 7,75% of substrate 7 (organic fertilizer)/ allow the combination of the substrates in order to adhere the Nitrate directive of the EU. The established differences in the active forms of nitrogen allow exploitation of programs for fertilizers with the participation of raw materials for biogas production. Our examination of substrates and show a content of heavy elements under the critical admissible limits. РЕЗЮМЕ: Направена е сравнителна екологична оценка на различни суровини за производство на биогаз. Оценката е на база качествата на различни субстрати: от енергийни култури (захарно, кръмно и червено цвекло, листна маса от Пауловния- пролетни и есенни листа, царевичен силаж и силаж Пауловния), три вида тор (говежда от безпостелно отглеждане, свинска и несменяема птича торова постеля), както и субстрати от отпадни води на ПС (първична,вторична и смесена утайка). Изследваните суровини бяха анализирани по следните показатели: Dry matter (DM) ,Organic matter (OM), Organic Carbon (OC), Nitrogen Kjeldahl (N), Nitrogen Ammonium (N -ammonium), Nitrogen nitrate (N- nitrate), Patassium (K), P (Phosphorus), pH ( H2O) and heavy metals – arsenic (As), Cadmium (Cd), Chromium ( Cr), Copper (Cu), Mercury (Hg), Nickel (Ni), Lead (Pd), Zinc (Zn). Резултатите от изследваните субстрати получени от различни суровини ( растителни, органични торове и отпадни води) ни дават основание да направим следните заключения: Изследваните субстрати по физикохимични показатели отговарят на всички технологични изисквания на суровини за производство на биогаз (СВ, С и съотношение С:N). Установените различия в съдържанието на азот е границите на 1,8% (субсрат - листна маса Пауловния –пролет) до 7,75% (при субстрат 7 - органична тор) позволяват комбиниране на субстратите с оглед спазване на Нитратната директива на ЕС. Съществуващите различия в подвижните форми на азота дават възможност за разработване на програми за торете с участие на различни суровини за производство на биогаз. Проучените от нас субстрати показват съдържание на тежки елементи под критично допустимите граници

Agroecologic valuation of organic waste in different technologies of storage

A study has been conducted, based on three types of fertilizers / cow, pig and bird manure/ and bioshlam /received from clearing station/ that have been storage in different technologies in order to make an agroecologic valuation. The different types of fertilizers and wastewater in different technologies of storage have been analyzed according to the following parameters: Dry matter (DM) ,Organic matter (OM), Organic Carbon (OC), Nitrogen Kjeldahl (N), Nitrogen Ammonium (N -ammonium), Nitrogen nitrate (N- nitrate), Sulfates (SO4) , Calcium Oxide (CaO), Magnesium Oxide (MgO), Patassium (K), P (Phosphorus), pH (H2O) and heavy metals – arsenic (As), Cadmium (Cd), Chromium (Cr), Copper (Cu), Mercury (Hg), Nickel (Ni), Lead (Pd), Zinc (Zn). The received results show: variation of the percentage of dry material /from 92,12% to 4,03%/ according to the technology of storage. The way of storage has no essential influence on the content of carbon in the various stable fertilizers, where pig manure has the highest value of nitrogen in all technologies of storage. Near to the recommendable optimal values of C:N is stayed/dry/ and fresh/hard/ in all three fertilizer types. The technology of storage by the different manure types has no influence on the content of heavy metal. Exception can be observed as far as the zinc content in stale /fluid/ pig manure is concerned. High content of dry material can be observed in bioshlam received from clearing station. In both ways of production of variation in the content of dry material the ratio between the biogenic macro elements C:N remains. The free nitrogen forms (N-ammonium and Nnitrate) in bioshlam are higher to other organic waste in both technologies of storage. No values over the critically admissible in bioshlam are observed in both technologies of production

Measuring extravascular lung water: animals and humans are not the same

The evolution of extravascular lung water (EVLW) monitoring is an important step forward in the hemodynamic assessment of critically ill patients

Increased Extravascular Lung Water Reduces the Efficacy of Alveolar Recruitment Maneuver in Acute Respiratory Distress Syndrome

Introduction. In acute respiratory distress syndrome (ARDS) the recruitment maneuver (RM) is used to reexpand atelectatic areas of the lungs aiming to improve arterial oxygenation. The goal of our paper was to evaluate the response to RM, as assessed by measurements of extravascular lung water index (EVLWI) in ARDS patients. Materials and Methods. Seventeen adult ARDS patients were enrolled into a prospective study. Patients received protective ventilation. The RM was performed by applying a continuous positive airway pressure of 40 cm H2O for 40 sec. The efficacy of the RM was assessed 5 min later. Patients were identified as responders if PaO2/FiO2 increased by >20% above the baseline. EVLWI was assessed by transpulmonary thermodilution before the RM, and patients were divided into groups of low EVLWI (<10 mL/kg) and high EVLWI (≥10 mL/kg). Results. EVLWI was increased in 12 patients. Following RM, PaO2/FiO2 increased by 33 (4–65) % in the patients with low EVLWI, whereas those in the high EVLWI group experienced a change by only −1((−13)–(+5)) % (P = 0.035). Conclusion. In ARDS, the response to a recruitment maneuver might be related to the severity of pulmonary edema. In patients with incresed EVLWI, the recruitment maneuver is less effective

Towards a FISH-based karyotype of Rosa L. (Rosaceae)

The genus Rosa Linnaeus, 1753 has important economic value in ornamental sector and many breeding activities are going on supported by molecular studies. However, the cytogenetic studies of rose specks are scarce and mainly focused on chromosome counting and chromosome morphology-based karyotyping. Due to the small size of the chromosomes and a high frequency of polyploidy in the genus, karyotyping is very challenging for rose species and requires FISH-based cytogenetic markers to be applied. Therefore, in this work the aim is to establish a FISH-based karyotype for Rosa wichurana (Crepin, 1888), a rose species with several benefits for advanced molecular cytogenetic studies of genus Rosa (Kirov et al. 2015a). It is shown that FISH signals from 5S, 45S and an Arabidopsis-type telomeric repeat are distributed on five (1, 2, 4, 5 and 7) of seven chromosome pairs. In addition, it is demonstrated that the interstitial telomeric repeat sequences (ITR) are located in the centromeric regions of four chromosome pairs. Using low hybridization stringency for ITR visualization, we showed that the number of ITR signals increases four times (1-4 signals). This study is the first to propose a FISH-based R. wichurana katyotype for the reliable identification of chromosomes. The possible origin of R wichurana ITR loci is discussed

Transcriptome Analysis of Targeted Mouse Mutations Reveals the Topography of Local Changes in Gene Expression.

The unintended consequences of gene targeting in mouse models have not been thoroughly studied and a more systematic analysis is needed to understand the frequency and characteristics of off-target effects. Using RNA-seq, we evaluated targeted and neighboring gene expression in tissues from 44 homozygous mutants compared with C57BL/6N control mice. Two allele types were evaluated: 15 targeted trap mutations (TRAP); and 29 deletion alleles (DEL), usually a deletion between the translational start and the 3' UTR. Both targeting strategies insert a bacterial beta-galactosidase reporter (LacZ) and a neomycin resistance selection cassette. Evaluating transcription of genes in +/- 500 kb of flanking DNA around the targeted gene, we found up-regulated genes more frequently around DEL compared with TRAP alleles, however the frequency of alleles with local down-regulated genes flanking DEL and TRAP targets was similar. Down-regulated genes around both DEL and TRAP targets were found at a higher frequency than expected from a genome-wide survey. However, only around DEL targets were up-regulated genes found with a significantly higher frequency compared with genome-wide sampling. Transcriptome analysis confirms targeting in 97% of DEL alleles, but in only 47% of TRAP alleles probably due to non-functional splice variants, and some splicing around the gene trap. Local effects on gene expression are likely due to a number of factors including compensatory regulation, loss or disruption of intragenic regulatory elements, the exogenous promoter in the neo selection cassette, removal of insulating DNA in the DEL mutants, and local silencing due to disruption of normal chromatin organization or presence of exogenous DNA. An understanding of local position effects is important for understanding and interpreting any phenotype attributed to targeted gene mutations, or to spontaneous indels

Reduction of sanitary indicator microorganisms in anaerobic digestion of poultry litter in combination with silages

ABSTRACT: In connection with experiments aimed at optimizing anaerobic digestion of organic fertilizers, microbiological tests were performed to assess the epizootiological safety of resulting finished product to fertilize the soil by quantities of sanitary indicator microorganisms. Mixtures of substrates were composed and tested in technological parameters established for other experiments: mesophilic temperature regime 33oC, dry matter content of 7%, residence time in the digester 15 days. The substrates tested were beetroot, sugar and salad beets, corn silage and leaves of Paulownia elongata, and silage combinations of these feeds in ratio 60:40 with mulch litter from broiler chickens. It was found that significant reduction of microorganisms was realized in the final products, which were assessed as safe for soil application. The ensiling of poultry litter in combinations of these feeds had the effect of reducing the amount of microorganisms in almost all examined groups therein. The most significant was the reduction in the quantity of sanitary revealing bacteria in the manure after its ensilage with autumn leaves of P. elongata and beetroot, and the least - with the participation of sugar beet. Microbial reduction was to the greatest extent in bioslime produced by anaerobic digestion of poultry litter ensiled with corn. Of the silages from different types of beets with poultry manure, during anaerobic digestion highest decontamination was achieved in this with sugar beets. After anaerobic degradation of the studied feed without poultry litter, at least microorganisms were found in bioslimes from the three types of beet, as well from the silage from P. elongata. РЕЗЮМЕ: Във връзка с експерименти, насочени към оптимизиране на анаеробното разграждане на органични торове, са извършени микробиологични изследвания за преценка на епизоотологичната безопасност на получените крайни продукти за наторяване на почви според количествата на санитарно индикаторните микроорганизми. Съставени са смеси от субстрати при установени при други експерименти технологични параметри: мезофилен температурен режим 33оС, съдържание на сухо вещество 7%, време на престой в биореактора 15 денонощия. Изпитани са захарно, кръмно и салатно цвекло, царевичен силаж и листа от Paulownia elongata, както и силажи от комбинации на тези фуражи в съотношение 60:40 с торова постеля от пилета-бройлери. Установено е, че се осъществява значително редуциране на микроорганизмите в крайните продукти, които се преценяват като безопасни за внасяне в почвите. В най-голяма степен микробната редукция е при биошлама, получен при анаеробното разграждане на комбинацията от торова постеля с царевичен силаж. От трите вида цвекло, силажирани с птичи тор, при анаеробно разграждане се постига най-висока деконтаминация с участие на захарното цвекло. При анаеробно разграждане на изследваните фуражи без птичи тор най-малко микроорганизми се установяват в биошламовете от трите вида цвекло, както и в силажа от P. elongata