Changes in neuronal DNA content variation in the human brain during aging

The human brain has been proposed to represent a genetic mosaic, containing a small but constant number of neurons with an amount of DNA exceeding the diploid level that appear to be generated through various chromosome segregation defects initially. While a portion of these cells apparently die during development, neurons with abnormal chromosomal copy number have been identified in the mature brain. This genomic alteration might to lead to chromosomal instability affecting neuronal viability and could thus contribute to age-related mental disorders. Changes in the frequency of neurons with such structural genomic variation in the adult and aging brain, however, are unknown. Here, we quantified the frequency of neurons with a more than diploid DNA content in the cerebral cortex of normal human brain and analyzed its changes between the fourth and ninth decades of life. We applied a protocol of slide-based cytometry optimized for DNA quantification of single identified neurons, which allowed to analyze the DNA content of about 500 000 neurons for each brain. On average, 11.5% of cortical neurons showed DNA content above the diploid level. The frequency of neurons with this genomic alteration was highest at younger age and declined with age. Our results indicate that the genomic variation associated with DNA content exceeding the diploid level might compromise viability of these neurons in the aging brain and might thus contribute to susceptibilities for age-related CNS disorders. Alternatively, a potential selection bias of "healthy aging brains" needs to be considered, assuming that DNA content variation above a certain threshold associates with Alzheimer's disease.     

海洋生境修复和生物资源养护原理与技术研究进展及展望

由于人类活动导致的海洋生境、生态系统以及生物资源的衰退已经引起了全球的高度重视。以生态修复的原理为基础,综述了海洋生境修复和生物资源养护的关键设施、关键技术、监测与效果评价以及综合管理的国内外研究进展,并对海洋生境修复与生物资源养护的研究热点和重点进行了展望,以期为我国海洋生境修复和生物资源养护提供参考。     

不同手术方式对腰椎间盘突出症治疗的效果比较分析

目的:探讨应用微创技术治疗腰椎间盘突出症的疗效.方法:对我院脊柱骨科自2005年l1月～2010年10月收治的128例腰椎间盘突出症患者应用不同手术方式进行治疗,其中应用APLD( automated percutaneous lumbar discectomy)治疗单纯腰椎间盘突出症48例;应用MED(microendoscopic discectomy)治疗复杂型腰椎间盘突出症42例;应用传统后路椎板开窗技术治疗单纯腰椎间盘突出症38例;比较各组手术时间、术中出血、术后住院时间、疗效及并发症.术后均随访8个月～3年,观察复发情况.结果:三组不同手术方式手术时间及优良率比较差异无统计学意义(P＞0.05),APLD及MED组术中出血量及术后住院时间与传统手术组比较,差异有显著性(P＜0.05);术后随访疼痛全部缓解,无复发.结论:在严格掌握适应症的基础上,采用微创技术和采用传统手术治疗腰椎间盘突出症疗效相当,但微创技术创伤小、术中出血量少,术后住院日短,恢复快,优于传统手术.     

Extraction of DNA from soil using nanoparticles by magnetic bioseparation

Aims: To develop a simple, rapid and inexpensive soil DNA extraction protocol. Methods and Results: The protocol relies on the use of superparamagnetic silica‐magnetite nanoparticles for the isolation and purification of DNA from soil samples. DNA suitable for use in molecular biology applications was obtained from a number of soil samples. Conclusions: The DNA extracted using the tested method successfully permitted the PCR amplification of a fragment of the bacterial 16S rDNA gene. The extracted DNA could also be restriction endonuclease digested. Significance and Impact of the Study: The protocol reported here is simple and permits rapid isolation of PCR‐ready soil DNA. The method requires only small quantities of soil sample, is scalable and suitable for automation.     

Diet of the Iberian hare (Lepus granatensis) in a mountain ecosystem

The diet of the Iberian hare (Lepus granatensis) was studied through microhistological pellet analysis in two areas from a mountain ecosystem in Central Portugal. Fecal pellets were collected monthly in 24 plots spatially distributed throughout the two study areas. For each period, a sample of 15 to 20 pellets was milled and 400 epidermal fragments were identified, by comparison with a reference collection. A wide range of plant species was observed in hare’s diet. Grasses represent the basis of the Iberian hare diet, with frequencies always higher than 50% in both study areas (annual average = 69.98%). Most of the 35 species of grasses assembled for the reference collection (91.43%) were identified in the pellets. Nevertheless, only six of these were consumed in proportions greater than 5%, being Anthoxanthum odoratum, Secale cereale and Agrostis spp. the species ingested in higher frequencies. The rate of grasses consumption reached 80.69% in winter but decreased in summer to around 55%. In this season, a concurrent rise in the ingestion of other plant groups, like herbs and shrubs, and of plant inflorescences was observed. This work provides the first results on the Iberian hare’s diet on mountain ecosystems, and suggests that the Iberian hare diet in a mountain ecosystem is similar to the observed in L. europaeus and L. timidus.     

梨形短梗蠕孢的分生孢子发育研究

采用玻片培养法对梨形短梗蠕孢(Trichocladium achrasporum)的分生孢子发育类型进行了研究。观察结果表明,梨形短梗蠕孢分生孢子既可以在菌丝顶端形成(顶生式),也可以在菌丝中间形成(间生式),其发育方式是全壁芽生式产孢。     

Fixation procedures for flow cytometric analysis of environmental bacteria

Analysis of environmental bacteria on the single cell level often requires fixation to store the cells and to keep them in a state as near life-like as possible. Fixation procedures should furthermore counteract the increase of autofluorescence, cell clogging, and distortion of surface characteristics. Additionally, they should meet the specific fixation demands of both aerobically and anaerobically grown bacteria. A fixation method was developed based on metal solutions in combination with sodium azide. The fixation efficiencies of aluminium, barium, bismuth, cobalt, molybdenum, nickel, and tungsten salts were evaluated by flow cytometric measurement of the DNA contents as a bacterial population/community stability marker. Statistical equivalence testing was involved to permit highly reliable flow cytometric pattern evaluation. Investigations were carried out with pure cultures representing environmentally important metabolic and respiratory pathways as controls and with activated sludge as an example for highly diverse bacterial communities. A mixture of 5 mM barium chloride and nickel chloride, each and 10% sodium azide was found to be a suitable fixative for all tested bacteria. The described method provided good sample stability for at least 9 days.     

A test on different aspects of diatom processing techniques

A factorial experiment was carried out in order to assess the effects of four treatment steps in diatom processing techniques (the quantity of hydrogen peroxide added to the sample, the time allowed to the digestion reaction, the use of centrifugation and the method used to settle the frustules onto the coverslip) on different aspects of the quality of the resulting microscopic slide: the concentration of organic and inorganic matter, the density, randomness and homogeneity of the frustule distribution in the slide, and the abundance of broken frustules and girdle views. The results show no significant effects of any of the experimental treatments on the proportion of broken frustules in the samples. No significant effects were related to the duration of the oxidation reaction in any of the aspects evaluated in the quality of the slides. A sample:hydrogen peroxide ratio of 1:1 for a 6-h oxidizing reaction can be used to optimize the protocol. Despite being more time consuming, the use of successive decantations instead of centrifuging is more appropriate to achieve better results. Slide quality can also be improved by leaving the coverslips to dry at room temperature in order to improve the distribution of frustules.     

Imprint cytology of vacuum-assisted breast biopsy specimens: a rapid diagnostic tool in non-palpable solid lesions

Objective:  Imprint cytology provides a rapid preliminary diagnosis shortly after the completion of breast biopsy. This study aims to assess the validity of imprint cytology for the pre-operative diagnosis of non-palpable mammographic solid lesions excised by vacuum-assisted breast biopsy (VABB).
Methods:  Seventy-two women with non-palpable Breast Imaging Reporting and Data System 3 and 4 mammographic solid lesions without microcalcifications underwent VABB on the stereotactic Fischer's table with 11-G Mammotome vacuum probes. Imprint samples were examined (Diff-Quick stain, modified Papanicolaou stain and May-Grünwald–Giemsa). The cores were dipped into a CytoRich Red Collection fluid for a few seconds in order to obtain samples with the use of the specimen wash. After the completion of cytological procedures, the core was prepared for routine pathological study. The pathologist was blind to the preliminary cytological results. The cytological and pathological diagnoses were comparatively evaluated.
Results:  The sensitivity of the cytological imprints for cancer was 90%. The specificity of the method for cancer diagnosis was 100%. Two precursor lesions were present in the material: one case of atypical ductal hyperplasia, which was successfully detected, and one case of lobular neoplasia, which escaped detection. The cytological imprints were inadequate in four out of 72 cases (5.6%), but none of them were included within the malignant subgroup.
Conclusions:  Imprint cytology seems to be an important adjunctive tool in the management of patients with non-palpable mammographic solid lesions. Its very satisfactory sensitivity and optimal specificity could establish its use in general clinical practice.