Quantitative variations of the isoforms in haptoglobin 1-2 and 2-2 individual phenotypes

Haptoglobin is a hemoglobin-binding protein presenting in humans three distinct phenotypes (Hpt 1-1, Hpt 1-2, or Hpt 2-2). The Hpt 1-2 and Hpt 2-2 phenotypes are in turn represented by populations of isoforms. The relative amounts of the major isoforms of Hpt 1-2 and Hpt 2-2 were found to differ not only in different individuals, but also in the same individual before and after a physical effort. Exercise-dependent changes in the plasma concentrations of ascorbate, urate, alpha-tocopherol, retinol, and glutathione were also observed, but correlations between such changes and those of the amount for any isoform were not found. Samples of Hpt 1-2 or Hpt 2-2 were challenged with oxidants (H(2)O(2) with ferrous ions, spermine-NO, KO(2), and 3-morpholinosydnonimine), but the isoform levels were not altered. Hpt 2-2 isoforms were present in Hpt 1-2, as minor species. Furthermore, different isoforms exhibited different hemoglobin binding abilities. Thus, these parameters should also be taken into consideration in studies correlating Hpt phenotypes prevalence with pathologies or functional differences.     

A modified protein assay from microgram to low nanogram levels in dilute samples

In this article, we present a modified and improved protein assay that was previously described as “amidoschwarz assay” by Schaffner and Weissmann [13]. Our improved protein assay is user-friendly and 30–40 times more sensitive than the earlier method. The assay was developed into three formats (macro-, micro-, and nanoassay) with trichloroacetic acid (TCA) as protein precipitating agent, measuring up to 96 samples. The macro and micro formats of this assay require a single reagent staining with amido black of protein dots bound to nitrocellulose membrane with lowest protein measurements to 1 and 0.1 μg, respectively. On the other hand, the nanoassay, with combination staining of amido black followed by colloidal gold, can extend the detection limit to 2.5 ng of protein. Protein concentrations were determined by densitometry and/or spectrophotometry. This assay is compatible with many ionic and non-ionic detergents. This improved protein assay provides an additional choice to researchers in measuring total protein concentration accurately in dilute biological samples as low as 0.125 μg/ml prior to their biochemical analysis such as in comparative proteomics.     

兰州市1907例不同年龄正常人群骨密度测定综合分析

目的：了解兰州地区正常人群骨密度的变化特点,分析其变化规律,为预防和治疗骨质疏松症提供科学依据。方法：使用天津圣鸿公司SHY-I数字式骨密度测定仪对兰州地区1907人进行检测,其中男1381例,女526例,分别做左前臂尺、桡骨测量。年龄20～85岁,每10岁为一年龄组进行统计分析。结果：男、女组骨密度峰值均在30-39岁,峰值后随年龄增加而骨密度下降,女性下降较男性显著。骨量减少及骨质疏松患病率在40岁后随年龄增长而增高,女性高于男性。老年人骨量减少及骨质疏松患病率高于中青年人,老年女性骨质疏松患病率与老年男性比较有明显差异（P〈0．05）。结论：兰州地区健康人群骨密度随年龄变化,并与性别有关。骨密度的检测在骨质疏松症的早期预防和治疗中具有重要意义。     

二龄草鱼脾脏、肝脏组织高表达甘露糖结合凝集素mRNA

Innate immunity is expected to be very important in fish. Mannose-bingding lectin (MBL) participates in the innate immune system as an activator of the complement system and as an opsonin after binding to certain carbohydrate structures on microorganisms. In this experiment, total mRNA was isolated from spleen, liver, gills, thymus, head kidney and kidney of adult and immature grass carp Ctenopharygodon idllus. The cDNA of MBL was obtained by RT-PCR using total mRNA from the spleen of carp as template. Such cDNA was labled with ^32p and used as probe for Northern analysis, and autoradiographic signals were quantified by densitometry analysis. The results showed that MBL was high expressed in the spleen and liver and low in gills, thymus, head kidney and kidney of adult grass carp, and MBL was much lower expressed in spleen and liver of immature grass carp than those of adult grass carp. The results might partially explain why immature grass carp are vulnerable to grass carp hemorrhage virus (GCHV) whereas adult grass carp are not.This suggested that MBL mav be an imoortant anti-GCHV factor [Acta Zoologica Sinica 50 (1): 137 - 140. 2004].     

In situ quantification of aberrant p53 in colorectal neoplasia

Aberrant p53 protein accumulation was measured immunohistologically in 342 colorectal paraffin-embedded tissue sections from 115 patients (24 with adenocarcinoma, 59 with adenoma and 32 'hospital controls'). Subjective scoring was compared with quantitative cell imaging, including dichotomous (p53⁺/p53^-) status, ng p53^mut mg^-1 enterocyte protein, and tumour burden and patient body 'burden' of aberrant p53. A total of 62.5% cancer patients, 23.7% adenoma patients and 3.1% hospital controls were accorded p53⁺ status on the basis of p53 quantification. Quantitative p53⁺/p53^- assignment had a stronger inverse association with survival (χ²=6.17, p=0.013, Kaplan-Meier test) than subjective 'visual estimation' (χ²=0.57, p=0.449). There was a strong inverse relationship between the p53 'body burden' and the months of post-diagnosis survival (hazard ratio=1.42, p=0.0004, Cox proportional hazards). Absolute quantification for inactivated p53 permits objective and reproducible scoring, adjusts for intra-laboratory immunostaining 'batch effects', corrects for fixation artefacts, and standardizes for inter-laboratory differences in fixation, antibody selection and staining method. Clinically, in situ quantification of p53 will permit more accurate survival prognoses and will inform therapy selection and dose. Ultimately, accurate quantitative tissue/blood p53 correlations may provide a minimally invasive and systemic surrogate measure for these same clinical purposes.     

Multivariate analysis of multispectral remote sensing data on grasslands from different soil types

Summary From an area of 60 sq. km in East-Flanders (Belgium), two sets of stereoscopic aerial photographs were taken. After selection of 13 soil types, 58 grasslands (mainly permanent ones) were localized. Data on species composition (S1 taxa) were quantitatively sampled with a double meter.Density values were obtained from both emulsions and using three filters. Using the densitometric varishles (extended to 22) or taxa variabies. Minimum Variance Clustering (MVC) and Principal Component Analysis (PCA) were applied. The recognition of different grasslandtypes and their relative position in both new sample point spaces was discussed as well as their relation to soil type. The results obtained by both Canonical Analyses (CCA and CVA) were examined and where necessary compared to the searce applications in vegetation ecology.Finally, the possible reasons for the meagre results were briefly discussed.Nomenelature follows De Langhe e.a. (1973).     

Microfiber reinforcement of an arthropod cuticle

Summary 1. According to its fine structure arthropod cuticle is a laminated composite material. The exoskeleton of a spider is surveyed in an electron microscopical study as to the characteristics of its fiber reinforcement.2. Five types of fiber arrangement are distinguished, ranging from unidirectional to continuously rotating fiber orientation and with three types combining elements of both of these extreme cases. The unidirectional type is rare and its occurrence confined to the innermost part of the walking leg endocuticle and to parts of articular membranes. Quite contrary a continuous and regular rotation of fiber direction is common. In hard cuticle (exocuticle) this type of reinforcement is found exclusively.3. Lamellar width, indicating a 180° rotation of the fiber direction, varies within wide limits. Variation is due to varying numbers of fiber layers and the amount of directional change between them (ca. 0.6–24°, type E).4. The diameter of the chitin/protein microfiber amounts to ca. 35 Å. Its length measured up to 1,5 m in the preparations. A periodicity of ca. 80 Å is noted. The precision of fiber alignment within the cuticle is high.5. In the cuticle of the opisthosoma and the articular membranes endocuticular lamellae are taking a wavy course, which is interpreted to be in relation to the cuticle's softness.6. The mechanical implications of both an increasing number of fiber directions in a composite material and of the different types of fiber reinforcement found in the spider cuticle are demonstrated by a calculation of the directional properties of the moduli of elasticity in tension and shear. The predictable strong influence of both the fiber content and the binder material's E-modulus on the mechanical properties of cuticle is illustrated.7. Instead of a trajectorial design a continuous rotation of the fiber orientation is found in all exocuticles. Thus the spider does not make use of the special advantage (high strength/weight ratio) of unidirectional fiber arrangement in places where it seemed most likely.Many thanks are due to Ms. H. Tscharntke for expert assistance with electron microscopy and photographic work. Dipl.-Ing. F. Och and Dr. G. Haberl (Messerschmitt-Bölkow-Blohm GmbH, Ottobrunn) provided the calculations for Figs. 14 and 15. Their generous help is gratefully acknowledged. Dr. K. Frahm (Universität München, Fakultät f. Tiermedizin) kindly shared his densitometer. Ms. F. Althaus carefully carried out the drawings.     

Towards quantitative cytotaxonomy of primates: Idiogram construction of great ape chromosomes by «fourier-warping»

To faciliate quatitative comparative cytogenetics of primate chromosomes we present a new type of idiogram from human and great ape chromosomes. These idiograms were obtained by computer densitometry and «Fourier-Warping» (FW) as described elsewhere (Maurier & Wienberg, 1991). In contrast to published idiograms, the «FW-idiograms» represent the mean position, width and density of each band and the mean resolution of the chromosome sample under investigation. In species that deverged about 6–12 my we were able to extract the representative banding features and to demonstrate conservation and change in the karyotype of human and great apes.     

Thin layer chromatographic analysis of glucose and maltose in estivated Biomphalaria glabrata snails and those infected with Schistosoma mansoni

Thin layer chromatography was used to analyze the glucose and maltose concentrations of the digestive gland–gonad complex (DGG) of uninfected-estivated Biomphalaria glabrata snails and estivated B. glabrata patently infected with Schistosoma mansoni. All snails were estivated in a most chamber at a relative humidity of 98 ± 1% and a temperature of 23 ± 1 °C for 14 days. Carbohydrates were extracted from the DGG with 70% aqueous ethanol, and extracts were analyzed on silica gel preadsorbent plates using ethyl acetate–glacial acetic acid–methanol–water (60:15:15:10) mobile phase, α-naphthol–sulfuric acid detection reagent, and quantification by densitometry. The concentrations of glucose and maltose were significantly reduced in both uninfected-estivated snails and infected-estivated snails.     

Morphometric and densitometric study of the biogenesis of electron-dense granules in Fonsecaea pedrosoi

Fonsecaea pedrosoi is a polymorphic pathogenic fungus, etiological agent of chromoblastomycosis, that synthesizes a melanin-like pigment. Although this pigment has been described as a component of the outer layers of the cell wall, electron-dense cytoplasmic bodies have also been visualized. In this work, we have correlated the appearance of intracellular electron-dense granules with the melanization process in F. pedrosoi. For this, conidial forms were grown under conditions where melanin was not synthesized. Afterwards, cells were incubated in Hank's medium supplemented with bovine fetal serum, at 37 degrees C, to stimulate the pigment production. The genesis of cytoplasmic bodies, with different stages of electron density, was demonstrated by transmission electron microscopy. The appearance of fungal acidic compartments, visualized by confocal laser scanning microscopy in cells stained with acridine orange, was time coincident with the formation of electron-dense granules observed by transmission electron microscopy. The quantification of granule numbers as well as morphometric and densitometric studies were performed.