Improvements in cytological preparations for fluorescent in situ hybridization in Passiflora

Cytological preparations for the fluorescent in situ hybridization (FISH) technique require cytoplasm-free metaphases, with well-spread chromosomes, for the localization of DNA sequences and chromosome mapping. We tested various procedures for FISH analysis of Passiflora cacaoensis, P. gardneri and hybrid F? progeny of P. gardneri x P. gibertii. Two treatments with four enzymes and three incubation times were compared. The material was treated with 1.0 M HCl before enzymatic digestion. The following criteria were used to determine the quality of the metaphases: a) lack or presence of cytoplasm; b) well-spread chromosomes or with overlap; c) complete or incomplete chromosome number (2n). The enzyme Pectinex(?) SP ULTRA gave the best performance, with the shortest incubation time. The best results were observed after 30 min of incubation; more than 70% of the metaphases did not have large amounts of cytoplasm or overlapping chromosomes, and about 75% maintained the chromosome number. FISH was carried out using a 45S rDNA probe (pTa71) labeled with biotin and detected with fluorescein isothiocyanate. Sites with strong staining and without nonspecific signals were observed. Our methodological adaptations allowed the preparation of metaphase slides of high quality for the FISH technique, with less time required for the preparation of samples.     

The role of SSDL in quality assurance in radiotherapy

This paper describes the role of the Polish Secondary Standard Dosimetry Laboratory (SSDL) in quality assurance in radiotherapy by means of providing calibration of ionisation chambers, TLD postal dosimetry audits and end-to-end audits for radiation therapy. A historical review of the methods and results are presented. The influence of the SSDL in Warsaw on radiation protection of patients in Poland is discussed. The International Atomic Energy Agency together with World Health Organisation (IAEA/WHO), through its network of SSDLs around the world, propagates newly developed methods for calibration and auditing. Suitable high quality equipment was provided by the IAEA, as well as special materials and technical support to the SSDL in Warsaw. The activity of the SSDL and the services provided for Polish radiotherapy centres have resulted in a reduction of discrepancies between planned doses and doses delivered to patients. The newly tested IAEA methods of end-to-end on-site dosimetry audits allow for monitoring and improving the quality of IMRT in Poland. The traceability of standards used for the calibration of therapy level dosimeters from Polish radiotherapy centres is assured by the IAEA dosimetry laboratory. The consistency of methods performed in the Polish SSDL with the ISO:17025 norm is supervised by the Polish Centre for Accreditation – a member of International Laboratory Accreditation Cooperation (ILAC), for calibration and testing. Due to the rapid technological development of radiotherapy, special attention has to be paid to newly developed methods for dosimetry auditing and institutions which provide services for assuring radiation safety of patients.     

Long-lasting inward current in snail neurons in barium solutions in voltage-clamp conditions

Summary The inward membrane current was recorded under voltage clamp from nonbursting neurons of the snailHelix pomatia in Na-free solutions containing Ba ions but no other divalent cations. The inward current was separated into two components: (i) an early fast inactivating component and (ii) a smaller long-lasting component. Both components were dependent on the external Ba concentration. It is concluded that both components of the inward current are carried by Ba ions. The activation of the early fast inactivating component of the inward current occurred at more positive membrane potential than that of the long-lasting component. The shape of the inactivation curve for the peak value of the inward current was similar to that for the long-lasting component. The potentials of half-inactivation for the peak value of the inward current and for its long-lasting component were –28 and –22 mV, respectively. The blocking effect of Co⁺⁺ on the early fast inactivating component was substantially greater. In some neurons after treatment with 15mm Co⁺⁺ only the long-lasting component was recorded. The activation kinetics of the long-lasting component of the inward current were analyzed using the Hodgkin-Huxley equations. The results could be explained by assuming that two components of the inward current in Na–Ca-free solution with Ba ions flowed through the two different channels. The significance of the long-lasting inward current for the normal spike generation is discussed.     

The role of leucine in ketogenesis in starved rats.

The quantitative significance of the conversion in vivo of L-[U-14C]leucine to ketone bodies was determined in rats starved for 3 or 48 h. In animals starved for 3 h, 4.4% of ketone-body carbon is derived from the metabolism of leucine, and in rats starved for 48 h the corresponding value is 2.3%. This conversion occurs rapidly, and the specific radioactivity of ketone bodies in blood is maximal at 2 min after the intravenous injection of labelled leucine for both periods of starvation. The flux of leucine in the blood is 1.01 and 1.04 mumol/min per 100 g body wt. respectively for animals starved for 3 and 48 h. The specific radioactivity of blood ketone bodies was compared at 2 min after the injection of labelled leucine, lysine and phenylalanine. The specific radioactivity was 4-5 fold higher with leucine than with lysine or phenylalanine.     

Twinning in thoroughbred mares in Poland

Of the thoroughbred population bred in Poland from 1952 to 1976, an average of 3.3% twinnings were registered. Of these averages 73% resulted in abortion, 11% in still births, and 16% in the live birth of one or both of the foals. Twins were not any more valuable for racing or breeding than normal. Mares mated in June had a twinning coefficient of 6.6% and those mated from February to May had one from 3.0% to 3.6%. The twinning coefficient was higher for mares which had not lactated during the previous year (6.0%, p 相似文献   

Warm ischemia-induced alterations in oxidative and inflammatory proteins in hepatic Kupffer cells in rats

The aim of the study was to investigate the impact of ischemia/reperfusion injury on the proteome of Kupffer cells. Lean Zucker rats (n = 6 each group) were randomized to 75 min of warm ischemia or sham operation. After reperfusion for 8 h, Kupffer cells were isolated by enzymatic perfusion and density gradient centrifugation. Proteins were tryptically digested into peptides and differentially labeled with iTRAQ (isobaric tags for relative and absolute quantitation) reagent. After fractionation by cation exchange chromatography, peptides were identified by mass spectrometry (ESI-LC-MS/MS). Spectra were interrogated against the Swiss-Prot database and quantified using ProteinProspector. The results for heat shock protein 70 and myeloperoxidase were validated by ELISA. Quantitative information for more than 1559 proteins was obtained. In the ischemia group proteins involved in inflammation were significantly up-regulated. The ratio for calgranulin B in the ischemia/sham group was 1.81 +/- 0.97 (p < 0.0001), for complement C3 the ratio was 1.81 +/- 0.49 (p < 0.0001), and for myeloperoxidase the ratio was 1.30 +/- 0.32. Myeloperoxidase was only recently documented in Kupffer cells. The antioxidative proteins Cu,Zn-superoxide dismutase (1.34 +/- 0.19; p < 0.001) and catalase (1.23 +/- 0.43; p < 0.001) were also elevated. In conclusion, ischemia/reperfusion injury induces alterations in the Kupffer cell proteome. Isotope ratio mass spectrometry is a powerful tool to investigate these reactions. The ability to simultaneously monitor several pathways involved in reperfusion stress may result in important mechanistic insight and possibly new treatment options.     

Control of Chagas disease in blood banks in Colombia

The screening programs for the Chagas disease agent, Trypanosoma cruzi, were examined in Colombian blood banks and, as a consequence, several procedural improvements in the blood bank network were recommended. Screening strategies and techniques were examined, as well as the action taken when seropositive donors were discovered. From a total of 180 blood banks in 33 departments, 103 banks in 20 departments answered the survey. The 103 banks collected 291, 105 units of blood, corresponding to 66.6% of all units collected in the country in 1997. Of these blood units, 99.6% were screened for Chagas trypanosomes; 3,321 (1.2%) of 287,048 were found positive for anti-T. cruzi. The data were grouped by department; geographical differences for seroprevalence rates varied markedly between 0% and 12.6%. The most commonly used serological technique was ELISA, but only 33.2% of the positive samples for anti-T. cruzi underwent further confirmatory testing, mainly through indirect immunofluorescent test. Most (95.1%) of the blood banks used basic, internal quality control procedures, and 73.8% sent positive samples to other laboratories for external quality control.     

Site-specific recombinases in genetic engineering: Modern in vivo technologies

A review was conducted over the current achievements in the area of site-specific recombinases (SSR) and their applications for manipulations with pro- and eukaryotic genomes. The principles of SSR functioning and the types of genetic rearrangements catalyzed by SSR were analyzed. The examples given in this review show the SSR potential to solve a wide range of basic and practical problems. To use different methods for solving these problems, however, would be more difficult or even impossible. The main directions for further developing the technology of site-specific recombination are the following: the use of SSR for a wider range of biological systems; generation of the SSR, which are characterized by strictly controlled expression in space and time; and the search for recombinases with new substrate specificity.     

广州地区农产品中有机磷农药残留调查

采用国家标准方法气相色谱(GC)法进行分析,通过对蔬菜、水果、粮食中16种有机磷农药残留量的检测,了解有机磷农药在广州地区农产品中的残留及其分布情况.50份农产品样品中有机磷农药各组分总的检出率为96.0%,其中蔬菜、水果、粮食检出率分别为96.8%、100%、80.0%.50份农产品样品中有机磷农药各组分总的超标率为36.0%,其中蔬菜、水果、粮食超标率分别为41.9%、21.4%、40.0%,16种有机磷农药中以马拉硫磷的超标率最高,为32.0%.不同组分有机磷农药在农产品中的残留量,以杀螟磷的平均含量最高,为1308.7μg·kg^-1,其次是辛硫磷和甲胺磷,分别为970.1μg·kg^-1和737.6μg·kg^-1;因此应提倡合理使用农药,加强农药使用的引导和监管.     

Trends in Risk Assessment of Chemicals in the European Union

Current legislation in the European Union (EU) requires a risk assessment for industrial chemicals. The underlying procedures and paradigms of such EU risk assessment for new and existing chemicals are explained. The risk assessment is performed according to a harmonised methodology, laid down in the Technical Guidance Documents (TGD). Important new, technical risk assessment aspects covered in a recent revision round of the TGD are highlighted. The most prominent change in the environmental TGD part is the addition of the marine risk assessment, including a Persistent Bioaccumulation and Toxicity (PBT) assessment. In the human health part a significant change is the new data requirement for reproductive toxicity. The performance of both the risk assessment and the risk reduction phase of EU existing chemicals have been evaluated. An important conclusion was that our a priori knowledge on possible risks of chemicals is poor. The European Commission has recently launched a proposal (REACH) for drastically changing the risk management process of industrial chemicals in the EU. Major changes are a shift in responsibility from authorities to industry (including downstream users) for the safe use of chemicals, an acceleration of data collection for ‘non-assessed’ chemicals, and an authorization step for substances of very high concern.     

Sex differentials in frailty in medieval England

In most modern populations, there are sex differentials in morbidity and mortality that favor women. This study addresses whether such female advantages existed to any appreciable degree in medieval Europe. The analyses presented here examine whether men and women with osteological stress markers faced the same risks of death in medieval London. The sample used for this study comes from the East Smithfield Black Death cemetery in London. The benefit of using this cemetery is that most, if not all, individuals interred in East Smithfield died from the same cause within a very short period of time. This allows for the analysis of the differences between men and women in the risks of mortality associated with osteological stress markers without the potential confounding effects of different causes of death. A sample of 299 adults (173 males, 126 females) from the East Smithfield cemetery was analyzed. The results indicate that the excess mortality associated with several osteological stress markers was higher for men than for women. This suggests that in this medieval population, previous physiological stress increased the risk of death for men during the Black Death to a greater extent than was true for women. Alternatively, the results might indicate that the Black Death discriminated less strongly between women with and without pre‐existing health conditions than was true for men. These results are examined in light of previous analyses of East Smithfield and what is known about diet and sexually mediated access to resources in medieval England. Am J Phys Anthropol 143:285–297, 2010. © 2010 Wiley‐Liss, Inc.     

Use of Ranks in Comparisons of Variability in the Non-Normal Populations

A method of analysis for comparing the variability of two samples drawn from two populations has been developed. The method is also suitable for the nonnumeric form of data. A test based on ordered observations for testing the null hypothesis of equality of two variances has been given. The test statistic is a function of the sum of ranks assigned to smaller size sample. Ranking procedure has been modified to depict the variability in the data by the sum of ranks. The null distribution of the test-statistic has been worked out for small samples and it turns out to be chi-square distribution for large samples. The analytical procedure has been explained by a numerical example on the productivity and production of rice and wheat in India from 1950–51 to 1983–84.     

Genome mapping in silver fox. Syntenic genes in Carnivora

Hamster X fox somatic cell hybrids segregating individual fox chromosomes in different combinations were used to assign seven structural loci to fox chromosomes. The gene for ME1 was mapped on the VFU1 chromosome, the genes for ADK and PP being located on the VFU4 chromosome. The gene for GSR was assigned to the VFU7 chromosome and the genes for MPI and COT1 were assigned to the VFU15 chromosome. Localization of these genes enhances the established fox genetic map and extends the known syntenic homologies between the fox and other mammalian. The comparison of data on gene mapping has provided basis for suggestion that there are significant differences in rates of karyotypic evolution in many mammalian taxa.     

Gene-Enzyme Relationships in Histidine Biosynthesis in Bacillus subtilis

Mutants for 9 of the 10 steps in histidine biosynthesis have been isolated and identified by enzyme assay. Each locus has been mapped in relation to the aro cluster and to other histidine loci by deoxyribonucleic acid-mediated transformation. The genes which code for enzymes 3, 6, and 8 of the pathway are linked to the aro cluster. A major histidine linkage group is composed of the genes which specify enzymes 1, 2, 5, 7, and 10. The locus which codes for step 9 of the pathway is unlinked to any other identified his loci. The major histidine cluster is loosely linked to cysB and is unlinked to any of the loci concerned with aromatic amino acid biosynthesis.     

Spatial distribution of fine-roots in boreal forests in eastern Sweden

Investigations were carried out in six forest types in areas surrounding two Swedish nuclear power plants (Forsmark and Laxemar). The aim of the investigation was to determine the spatial distribution of fine-root biomass (live), necromass (dead) and standing crop (live + dead) and to test the use of the live/dead ratio as a vitality criterion. Soil cores were taken to depths with insignificant amounts of roots. The total amount of fine-root biomass (<1 mm in diameter) of tree species in the soil profile was 267, 317 and 235 g m^?2 for the Forsmark and 137, 371 and 50 g m^?2 for the Laxemar sites. The related necromass was 119, 226 and 184 g m^?2 and 87, 245 and 271 g m^?2. The biomass in the humus layer was 47, 7 and 48% for the Forsmark and 34, 26 and 7% for the Laxemar sites, as a percentage of the total live + dead fine roots in the soil profile. The related necromass in the humus layer was 13, 2 and 30% for the Forsmark and 13, 2 and 28% for the Laxemar sites. The live/dead ratio decreased with depth for both tree— and field-layer species and seems to be a most powerful vitality criterion of fine roots.     

Survey of strontium in mineral waters sold in Japan

The concentrations of strontium, calcium, and magnesium in 33 brands of natural mineral waters commercially available in Japan were determined by inductively coupled plasma-atomic emission spectrometry. The geometric mean values were 94.4 microg/L for strontium, 19.1 mg/L for calcium, and 2.82 mg/L for magnesium. Wide confidence intervals of 1.96-4539 microg/L for strontium, 0.865-421 mg/L for calcium, and 0.064-123 mg/L for magnesium were observed. The significant linear relationships among the three elements over a wide distribution range suggest that the synchronized variations of these elements are regulated by the natural ecosystem and not from accidental contamination from human activities or exceptionally high natural sources. Using the results of multiple linear regression analysis, the strontium concentration can be predicted by that of calcium with the appropriate power function. The results of this study suggest that mineral water can be an important nutritional source of strontium. As trace elements imbalance is often found in older patients with chronic renal failure, we propose that close attention of trace elements intake from trendy foods or beverages is necessary to prevent this hidden problem of a rapidly aging society.     

短时高温对加州新小绥螨发育的影响

为了探究高温逆境条件下加州新小绥螨的生存特性对其种群发育的影响,利用短时高温刺激试验,研究加州新小绥螨卵、幼螨、雌成螨在35、38、42、45 ℃等4个温度下、处理1～8 h后的孵化率、存活率、后期发育历期及繁殖的影响.结果表明: 处理温度越高,时间越长,卵和幼螨的存活率越低,其后续发育历期随处理温度升高和时间增加先缩短后延长.当温度为38 ℃,处理8 h后,卵的后续发育历期最短,为4.1 d.卵在45 ℃下处理2 h以上将不能正常孵化,而幼螨在45 ℃下处理8 h后不能存活;雌成螨产卵期和产卵量基本随温度升高先增加后降低,35 ℃处理8 h后,单雌产卵量最高,为38.9粒;38 ℃处理8 h后,单雌产卵量为36.7粒;45 ℃处理8 h后,单雌产卵量仅为14.5粒.短时高温主要影响加州新小绥螨的孵化率、存活率和后续发育历期,对雌成螨的产卵前期和存活率影响较小.     

Time trends in testicular cancer in Croatia 1983-2007: rapid increases in incidence, no declines in mortality

Testicular cancer, although a rare malignancy, represents the most common cancer in young male populations of Western origin. While increasing incidence trends of testicular cancer have been reported, mortality is declining in many high-resource settings. Using national data from the Croatian National Cancer Registry for the period 1983-2007, time trends were analysed by joinpoint regression and Age-Period-Cohort models. The present study is the first to analyse the testicular cancer trends in the Croatian population. Over the 25-year period, a mean number of 89 incident cases and 13 deaths were reported annually. The observed mean annual increases in age-standardised rates were 7.0% for incidence and 1.6% for mortality, with no abrupt linear changes (joinpoints) identified. The incidence rates of testicular cancer incidence have been steeply increasing in successive cohorts born since the mid-1930s. The rapid rise in testicular cancer incidence in the Croatian population appears to be one of the highest rates of increase recorded in Europe and worldwide. The lack of decline in the mortality rates over time, while based on relatively few deaths, highlights a need for improvements in diagnostics and management of therapy in Croatia in order to improve the survival and quality-of-life of testicular cancer patients.     

Olfaction in Drosophila

The fruit fly, Drosophila melanogaster, is equipped with a sophisticated olfactory sensory system that permits it to recognize and discriminate hundreds of discrete odorants. The perception of these odorants is essential for the animal to identify relevant food sources and suitable sites for egg-laying. Advances in the last year have begun to define the molecular basis of this insect's discriminatory power. The identification of a large multi-gene family of candidate Drosophila odorant receptors suggests that, as in other animals, a multitude of distinct odorants is recognized by a diversity of ligand-binding receptors. How olfactory signals are transduced and interpreted by the brain remains an important question for future analysis. The availability of genetic tools and a complete genome sequence makes Drosophila a particularly attractive organism for studying the molecular basis of olfaction.