NOR expression increases in interphase lymphocytes of Down syndrome babies/children as AgNORs surface, according to the mitogen concentration in the culture medium

The extra chromosome 21 of Down syndrome (DS) or trisomy 21 patients contains an average of 40 extra copies of rRNA genes and the in vivo/in vitro regulation of the activity of these genes is not fully understood. The objective of this work was to compare the NORs expression pattern in interphase lymphocytes of DS patients with regular trisomy 21 and control individuals according to phytohemagglutinin (PHA) concentration (0.37, 0.75, 1.48 and 2.21 ml) per 100 ml of medium. Because the AgNOR staining is an indicator of the active rRNA genes, comparison of the image analysis values of the AgNOR area in 72 h cultivated lymphocytes for each concentration of PHA between DS patients (N=30) and controls (N=24) provided a plausible conclusion on the regulation of the extra rRNA genes in DS lymphocytes. The nucleolus organizer regions area/total nuclear area (NORa/TNa) was calculated using an in-house computer program. Fifty consecutive interphases per PHA concentration were analysed for each individual, for determination of the NORa/TNa. In contrast to healthy controls, NORa/TNa of lymphocytes from DS patient babies/children (0-8 years old) increased gradually in parallel with the PHA concentration in the culture medium: 10.44+/-1.72% for 0.37 ml of PHA, 11.74+/-1.93% for 0.75 ml of PHA, 13.25+/-2.03% for 1.48 ml of PHA and 13.43+/-2.08% for 2.21 ml of PHA per 100 ml of medium. Contrary to control cells (in which the NORa/TNa ratio according to PHA concentration in the culture medium remains constant), DS interphase lymphocytes in culture do not down-regulate their NOR expression. These results obtained from interphase NORs are consistent with the previous results obtained by evaluating the mean of AgNOR+ chromosome number in metaphase cells, also in relation to the mitogen concentration in the culture medium.     

AgNOR increase in buccal epithelial cells of trisomy 21 infants

The aim of this study is to compare the Argilophilic Nucleolus Organizer Regions (AgNORs) level between Down syndrome (DS) patients and controls in a tissue sharing the same embryonic origin with the central nervous system and compare the results with those obtained recently by us from DS's lymphocytes. For this, buccal desquamating epithelial cells well known as the ectodermic origin were used. Since the AgNOR staining intensity is an indicator of the ribosomes biosynthesis rate, comparison of the image analysis values of the AgNOR area/total nuclear area (NORa/TNa) in buccal desquamating epithelial cells of DS patients and controls provided a plausible conclusion about the regulation/deregulation of the rRNA genes (rDNA) in these cells of DS babies/infants. The (NORa/TNa) proportion was calculated using an in-house computer program. Fifty buccal desquamating cells were analysed for each individual to determine the average NORa/TNa value per individual. In contrast to healthy controls, NORa/TNa proportion value of buccal epithelial cells from DS patients found significantly higher than that of the controls: (4.08 ± 1.16)% and (2.13 ± 0.55)%, respectively. This 92% increase is far higher than the expected value due to the extra rRNA genes on the extra-chromosome 21. Finally DS babies/infants exhibit very higher AgNOR expression increase in their buccal epithelial cells compared to controls. This is the first study that is available on the comparison of AgNOR expression levels in buccal epithelial cells between DS infants and their controls.     

AgNOR staining and quantification

Nucleolar organiser regions (NORs) are defined as nucleolar components containing a set of argyrophilic proteins, which are selectively stained by silver methods. After silver-staining, the NORs can be easily identified as black dots exclusively localised throughout the nucleolar area, and are called "AgNORs". The NORs' argyrophilia is due to a group of nucleolar proteins, which have a high affinity for silver (AgNOR proteins). A number of studies carried out in different tumour types demonstrated that malignant cells frequently present a greater AgNOR protein amount than corresponding non-malignant cells. Moreover, in cancer tissues AgNOR protein expression was found to be strictly related to the cell duplication rate. Over the past 12 years, the "AgNOR method" has been applied in tumour pathology for both diagnostic and prognostic purposes. However, the lack of a standardised silver-staining protocol has led to much misinterpretation of actual structures evaluated in individual studies. Indeed, the absolute AgNOR scores reported by different authors for the same types of tumour are scarcely comparable and the results produced by these investigations sometimes seem to be conflicting. In order to achieve definitive standardisation of the AgNOR method and produce comparable data in all laboratories, the "International Committee on AgNOR Quantitation" was founded, and during the first Workshop "AgNORs in Oncology" held in Berlin in 1993 guidelines for AgNOR protein evaluation were first defined. The present paper discusses the main technical aspects of NOR silver-staining, and critically evaluates the methods commonly employed for AgNOR protein quantification in routine cyto-histopathology.     

Prognostic relevance of AgNORs in tumor pathology

The importance of the analysis of the silver-stained nucleolar organizer regions (AgNORs) for prognostic purposes in tumor pathology has been reviewed. Current available data from the literature demonstrate that the evaluation of the quantity of interphase AgNORs is an independent prognostic factor in several types of human tumors. Results of our investigations indicate that AgNORs are the most powerful variable predicting survival in patients with pharyngeal carcinoma, multiple myeloma, male breast and prostate carcinoma. The combination of AgNOR counts and histologic pattern allows the stratification of patients with multiple myeloma, pharyngeal and prostate carcinoma into low- and high-risk groups, which could benefit from different therapy. Moreover, AgNOR analysis predicts response to treatment in adult patients with acute myelogenous leukemia, and appears as an independent prognostic factor in a prospective study on renal cell carcinoma. Therefore, AgNOR analysis is a really important prognostic factor for several human neoplasias. The experimental and theoretical justifications for AgNORs as a prognostic factor are also reviewed, in particular the strict correlation between AgNOR quantity and tumor cell doubling time. Lastly, the lack of prognostic significance of AgNOR analysis in some circumstances is critically discussed.     

Two-dimensional agnor evaluation as a prognostic variable in urinary bladder carcinoma: a different approach via total agnor area/nucleus area per cell

Traditional criterions are not sufficient to predict accurately the recurrence of transitional cell carcinoma of the urinary bladder. Therefore, we aimed to evaluate the AgNORs via total AgNOR area/nucleus area (TAA/NA) for each cell as a prognostic parameter, in TCC of urinary bladder. Tumor tissues of 20 consecutive cases of male bladder cancer patients were divided into two groups as middle differentiated (LG) and high grade (HG). The extra-tumoral tissue (ETT) samples of 10 males served as control group. A second control group (HC) consisted of five healthy and normal bladder tissue samples. The 3 microm of sections from each paraffin embedded tumoral, extra-tumoral and normal tissue samples served as patient and control groups. After deparaffinization and rehydratation steps, silver (AgNO(3)) staining of nucleolar organizer regions-associated proteins (AgNORs) was performed. Instead of Giemsa stain, we used Hematoxylin for contra staining. The images of the 100 analyzable nuclei from each tissue sample, transferred by means of a video camera and video capture card from microscope and recorded onto a computer. Software was prepared in Delphi language for analysis. Mean (E+02) TAA/NA values of HC, ETT, LG and HG groups were 6.97+2.80, 5.70+1.82, 7.80+3.22 and 9.24+3.88, respectively. Statistical comparisons have shown significant differences between all groups.In conclusion, mean TAA/NA per cell has a potential to be a prognostic parameter. Therefore, further evaluation of big patient series will be useful.     

Study of the nucleolar cycle and ribosomal RNA distribution during meiosis in triatomines (Triatominae, Heteroptera)

Aspects of nucleolar activity during spermatogenesis were assessed in three triatomine species (Panstrongylus megistus, Rhodnius pallescens and Triatoma infestans) using cytochemical and fluorescent staining techniques. Toluidine blue and a variant of critical electrolytic concentration (CEC) allowed the discrimination of rRNA providing structural details of the nucleolus and RNA distribution during meiotic cell division. Acridine orange fluorochrome staining permitted the differentiation of nucleic acids, and silver-ion impregnation made possible the observation of pre-nucleolar bodies (PNBs). Our results support the phenomenon known as "persistence of the nucleolar material", and the hypothesis of post-meiotic reactivation of rRNA genes. Nucleolar organizer regions (NORs) were observed in some metaphasic spermatogonial chromosomes in P. megistus and T. infestans. In P. megistus at diplotene-diakinesis, NORs were also detected in one of the sex chromosomes and in an autosome. Therefore, it may be inferred that, in triatomines, the nucleolus does not completely disappear, but persists in the form of small bodies that get together to form the next nucleolar cycle which, in the case of meiosis, will be completed if fertilization occurs and a new zygote is formed.     

AgNORs in breast tumours

There is evidence that the quantitative distribution of AgNOR proteins is a proliferation-related parameter that can be used as a prognostic index in tumour pathology. In breast cancer, some authors found a significant prognostic correlation of AgNOR protein quantity, whereas other did not. However, in all the reports dealing with AgNOR area (as opposed to count) this parameter was always turned out to be an independent prognostic indicator. The present study tests the significance of AgNOR proteins in a large series of primary breast carcinomas, exploring the associations between the AgNOR protein amount, as evaluated by image cytometry, and the other well-established prognostic markers commonly considered for breast cancer, along with patients' survival. Our results demonstrated a highly significant association between AgNOR protein quantity and tumour prognosis. Moreover, when the AgNOR area values were entered in multivariate analysis together with the other predictive parameters commonly considered in breast carcinomas, they showed an independent prognostic value together with Ki67-labelling index (LI), N-status and tumour size. Considering node-negative and -positive cases separately, the AgNOR protein area and Ki67-LI both come out as a independent predictors only in the latter group: the short follow-up time of our series (36 months median) could be responsible for this discrepancy.     

MR-visible brain water content in human acute stroke

Quantification of metabolite concentrations by proton magnetic resonance spectroscopy (1H-MRS) in the human brain using water as an internal standard is based on the assumption that water content does not change significantly in pathologic brain tissue. To test this, we used 1H-MRS to estimate brain water content during the course of cerebral infarction. Measurements were performed serially in the acute, subacute, and chronic phase of infarction. Fourteen patients with acute cerebral infarction were examined as well as 9 healthy controls. To correlate with regional cerebral blood flow (rCBF) SPECT-scanning using 99mTc-HMPAO as flow tracer was performed in the patients. Mean water content (SD) in the infarct area was 37.7 (5.1); 41.8 (4.8); 35.2 (5.4); and 39.3 (5.1) mol x [kg wet weight](-1) at 0-3; 4-7; 8-21; and >180 days after stroke, respectively. Water content increased between Day 0-3 and Day 4-7 (p = 0.034) and decreased from Day 0-3 to Day 8-21 (p = 0.028). Water content at Day 4-7 was significantly higher than in controls (p < or = 0.05). At the same time intervals, mean rCBF (SD) was 76 (23); 94 (31); 106 (35); and 64 (26)%, respectively. There was a significant increase in rCBF from Day 0-3 to Day 4-7 (p = 0.050) and from Day 0-3 to Day 8-21 (p = 0.028). No correlation between rCBF and water content was found. Water content in ischemic brain tissue increased significantly between Day 4-7 after stroke. This should be considered when performing quantitative 1H-MRS using water as an internal standard in stroke patients.     

Degradation pathways of dissolved carbon in landfill leachate traced with compound-specific (13)C analysis of DOC

The isotopic compositions of carbon compounds in landfill leachate provide insights into the biodegradation pathways that dominate the different stages of waste decomposition. In this study, the carbon geochemistry of different carbon pools, environmental stable isotopes and compound-specific isotope analysis (CSIA) of leachate dissolved organic carbon (DOC) fractions and gases show distinctions in leachate biogeochemistry and methane production between the young area of active waste emplacement and the old area of historical emplacement at the Trail Road Landfill (TRL). The active area leachate has low DOC concentrations (<200 mg l(-1)) dominated by fulvic acid (FA=160 mg l(-1)), and produces CH(4) dominantly by CO(2) reduction (D- excess=20.6 per thousand). Leachate generated in the area of older waste has high DOC (>4770 mg l(-1)) dominated by FA (4482 mg l(-1)) and simple fatty acids (acetic=1008 mg l(-1) and propionic=608 mg l(-1)), and produces CH(4) by the acetate fermentation pathway (D- excess=9.8 per thousand). CSIA shows an advanced degradation and a progressive accumulation of (13)C of fatty acids in leachate from the older area. The enriched (13)C value of FA (-20 and-26 per thousand for the older and active parts, respectively,) and of low molecular weight DOC (-8 and-27 per thousand) as well as of the bulk DOC (-21 and-25 per thousand) shows more advanced degradation in the older part of the landfill, which is consistent with the shift in the humic/FA ratios (0.05 and 0.18). The (13)C enrichment of acetate (-12 per thousand) above the (13)C of DOC (-21 per thousand) and of propionic acid (-19 per thousand), in older leachate, suggests that this acetate has not evolved from the simple degradation of larger organic molecules, but by homoacetogenesis from the enriched dissolved inorganic carbon (DIC) pool (8 per thousand) and H(2,) which produce a more enriched (13)C of acetate. In contrast, the (13)C of the minor acetate in the active area (-17 per thousand) indicates that CO(2)-reducing bacteria must be the primary consumers of H(2), which has resulted in enriched (13)C(DIC) (10 per thousand) and depleted (13)C(CH4) (-58 per thousand).     

Software compensation of eddy current fields in multislice high order dynamic shimming

Dynamic B(0) shimming (DS) can produce better field homogeneity than static global shimming by dynamically updating slicewise shim values in a multislice acquisition. The performance of DS however is limited by eddy current fields produced by the switching of 2nd and 3rd order unshielded shims. In this work, we present a novel method of eddy field compensation (EFC) applied to higher order shim induced eddy current fields in multislice DS. This method does not require shim shielding, extra hardware for eddy current compensation or subject specific prescanning. The interactions between shim harmonics are modeled assuming steady state of the medium and long time constant, cross and self term eddy fields in a DS experiment and 'correction factors' characterizing the entire set of shim interactions are derived. The correction factors for a given time between shim switches are shown to be invariable with object scanned, shim switching pattern and actual shim values, allowing for their generalized prospective use. Phantom and human head, 2nd and 3rd order DS experiments performed without any hardware eddy current compensation using the technique show large reductions in field gradients and offsets leading to significant improvements in image quality. This method holds promise as an alternative to expensive hardware based eddy current compensation required in 2nd and 3rd order DS.     

土壤Cd含量实验室与野外DS光谱联合反演

土壤重金属高光谱遥感建模理论上能够大大降低传统化学分析测定所需成本,正逐步发展为有效探查土壤污染空间分布与开展污染土壤综合防治的关键技术。然而土壤重金属高光谱遥感调查技术目前多局限于稳定可控条件下的实验室光谱模型,野外诸多因素（光照、湿度、土壤粗糙度等）影响下野外原位光谱模型的有效性已成为困扰该项技术大范围推广亟待突破的关键科学问题。以湖南衡阳市某矿区为例,分别利用ASD地物光谱仪和等离子发射光谱法测定46个土壤样品350~2 500 nm的实验室光谱和Cd含量,并在土壤取样时同步测量样品野外原位光谱。在运用DS（direct standardization）转换算法处理野外光谱的基础上,融合实验室光谱先验知识,基于主成分逐步回归建模方法开展了土壤Cd含量实验室与野外原位DS光谱联合反演实验,交叉验证了模型的稳定性。同时为深入探究实验室与野外原位DS光谱联合反演模型的有效性,将其与基于实验室光谱、野外原位光谱、野外原位DS光谱、实验室与野外原位光谱联合建立的主成分逐步回归模型开展了对比分析。结果表明：野外原位光谱反演模型精度（R2=0.56）明显低于实验室光谱反演模型（R2=0.64）,野外原位DS光谱反演模型与之相比精度有所提升（R2=0.66）;在野外原位光谱DS转换校正基础上,联合实验室光谱先验知识的土壤Cd含量反演模型精度最高,R2可达0.72。与此同时,实验室与野外原位DS光谱联合反演模型揭示482,565,979和2 206 nm波段对研究区土壤Cd含量有较好指示性,此结果与实验室光谱反演模型所识别的特征波段一致,两者物理意义相同。研究结果证实了实验室光谱先验知识以及DS转换算法能够提升野外原位光谱模型的可靠性,可为发展土壤Cd含量野外原位高光谱遥感探测提供重要的提供理论与方法支撑。     

PHASE TRANSITIONS IN Bi2Sr2(Cu1-zFez)O6+δ

We have investigated, by X-ray diffraction, a series of single crystals of Bi-based oxides with the nominal composition Bi₂Sr₂(Cu_1-zFe_z)O_6+δ(0≤z≤0.55). In this system we observed two structural phase transitions with the increase of the doping content. The first transition, from an incommensurate monoclinic phase to an incommensurate orthorhombic phase, occurs at a doping content of iron z_Fe=0.027. The second one corresponds to a phase transition from an incommensurate orthorhombic phase to a commensurate orthorhombic phase at z_Fe=0.34. The comparison of these results with those for more limited substitutions of Zn and Ni indicates the significant role of the insertion of the extra oxygen in the (Bi-O)_∞ double layers.     

Three-dimensional modeling of genome macroarchitecture on the basis of its structural changes after the action of radiation

At present, after 120 years of theoretical and experimental studies, the problem of the genome macroarchitecture as the highest level of interphase chromosome organization in the nucleus of somatic cells is still open. The problem of spatial organization of interphase chromosomes in the haploid nucleus of germ cells has never been studied. The three-dimensional modeling of spatial organization of part of the haploid genome (the second chromosome) in Drosophila melanogaster mature sperms is performed using mathematical methods and the methods of visualization of macromolecular biostructures. The frequency and arrangement of inversion breaks for 72 structural vg mutants were used as genetic markers under the assumption that both ends of each inversion are brought together and form loops of an appropriate size. For taking into account the spatial proximity and visualization of loop structures of the chromosome, modern methods of three-dimensional modeling with application of splines, Open GL library, Delphi, and Gmax were used. According to the model developed, the whole second chromosome in the nucleus of mature sperms is probably arranged in the form of the megarosette-loop structure, which can be assumed to be the fundamental ordered form of the genome macroarchitecture in haploid germs of higher organisms.     

Discovery of doubly magic 48Ni

In an experiment at the SISSI/LISE3 facility of GANIL, we used the projectile fragmentation of a primary 58Ni26+ beam at 74.5 MeV/nucleon with an average current of 3 &mgr;A on a natural nickel target to produce very neutron-deficient isotopes. In a 10-day experiment, 287 42Cr isotopes, 53 45Fe isotopes, 106 49Ni isotopes, and 4 48Ni isotopes were unambiguously identified. The doubly magic nucleus 48Ni, observed for the first time, is the most proton-rich isotope ever identified with an isospin projection T(z) = -4. It is probably the last doubly magic nucleus with "classical" shell closures accessible for present-day facilities. Its observation allows us to deduce a lower limit for the half-life of 48Ni of 0.5 &mgr;s.     

Organization of chromatin in the interphase mammalian cell

The use of imaging techniques has become an essential tool in cell biology. In particular, advances in fluorescence microscopy and conventional transmission electron microscopy have had a major impact on our understanding of chromatin structure and function. In this review we attempt to chart the conceptual evolution of models describing the organization and function of chromatin in higher eukaryotic cells, in parallel with the advances in light and electron microscopy over the past 50 years. In the last decade alone, the application of energy filtered transmission electron microscopy (EFTEM), also referred to as electron spectroscopic imaging (ESI), has provided many new insights into the organization of chromatin in the interphase nucleus. Based on ESI imaging of chromatin in situ, we propose a 'lattice' model for the organization of chromatin in interphase cells. In this model, the chromatin fibers of 10 and 30nm diameter observed by ESI, produce a meshwork that accommodates an extensive and distributed interchromosomal (IC) space devoid of chromatin. The functional implications of this model for nuclear activity are discussed.     

Observational constraints on the accelerating universe in the framework of a 5D bounce cosmological model

In the framework of a five-dimensional (5D) bounce cosmological model, a useful function f(z) is obtained by giving a concrete expression of deceleration parameter q(z)=q₁+{q₂}/{1+ln (1+ z)}. Then using the obtained Hubble parameter H(z) according to the function f(z), we constrain the accelerating universe from recent cosmic observations: the 192 ESSENCE SNe Ia and the 9 observational H(z) data. The best fitting values of transition redshift z_T and current deceleration parameter q₀ are given as z_T= 0.65_-0.12^0.25 and q₀ = - 0.76_-0.15^+0.15 (1σ). Furthermore, in the 5D bounce model it can be seen that the evolution of equation of state (EOS) for dark energy w_de can cross over -1 at about z=0.23 and the current value w_0de= - 1.15<- 1. On the other hand, by giving a concrete expression of model-independent EOS of dark energy w_de, in the 5D bounce model we obtain the best fitting values z_T= 0.66_0.08^+0.11 and q₀ = - 0.69_0.10^+0.10 (1σ) from the recently observed data: the 192 ESSENCE SNe Ia, the observational H(z) data, the 3-year Wilkinson Microwave Anisotropy Probe (WMAP), the Sloan Digital Sky Survey (SDSS) baryon acoustic peak and the x-ray gas mass fraction in clusters.     

Measurement of the Michel parameter rho in muon decay

The TWIST Collaboration has measured the Michel parameter rho in normal muon decay, mu(+)--> e(+)nu(e)nu (mu). In the standard model, rho = 3/4. Deviations from this value imply mixing of left- and right-handed muon and electron couplings. We find rho=0.750 80+/-0.000 32(stat) +/- 0.000 97(syst) +/- 0.000 23, where the last uncertainty represents the dependence of rho on the Michel parameter eta. This result sets new limits on the W(L)-W(R) mixing angle in left-right symmetric models.