Depicting the Spatial Distribution of Proteins in Human Tumor Tissue Combining SELDI and MALDI Imaging and Immunohistochemistry

Carcinoma tissue consists of not only tumor cells but also fibroblasts, endothelial cells or vascular structures, and inflammatory cells forming the supportive tumor stroma. Therefore, the spatial distribution of proteins that promote growth and proliferation in these complex functional units is of high interest. Matrix-assisted laser desorption/ionization imaging mass spectrometry is a newly developed technique that generates spatially resolved profiles of protein signals directly from thin tissue sections. Surface-enhanced laser desorption/ionization mass spectrometry (MS)combined with tissue microdissection allows analysis of defined parts of the tissue with a higher sensitivity and a broader mass range. Nevertheless, both MS-based techniques have a limited spatial resolution. IHC is a technique that allows a resolution down to the subcellular level. However, the detection and measurement of a specific protein expression level is possible only by semiquantitative methods. Moreover, prior knowledge about the identity of the proteins of interest is necessary. In this study, we combined all three techniques to gain highest spatial resolution, sensitivity, and quantitative information. We used frozen tissue from head and neck tumors and chose two exemplary proteins (HNP1–3 and S100A8) to highlight the advantages and disadvantages of each technique. It could be shown that the combination of these three techniques results in congruent but also synergetic data. (J Histochem Cytochem 58:929–937, 2010)     

Endomycopsis montevidensis n. sp.

A new species of the genusEndomycopsis is proposed. It was isolated from waste water, at Montevideo-Uruguay.By its morphological and biochemical proprieties it differs from allEndomycopsis previously described.

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Este trabajo fue desarrollado en el Laboratorio de Micologia de la Facultad de Química de Montevideo, Uruguay y en uso de una beca de la Universidad Federal de Pernambuco para perfecciobamiento en el estudio de las levaduras.     

INTERVENTION OF BRAIN CORTEX PHOSPHOLIPIDS IN PYRIDOXAL PHOSPHATE-DEPENDENT REACTIONS

We have examined the role of brain cortex phospholipids in regulating some enzymic reactions specific to the CNS. The study was carried out at the level of the reactions concerned with GABA formation, both in vivo and in vitro, and included investigation of the specificity of the effect. It was found that the brain cortex phospholipids enhance the transformation of exogenous vitamin B₆ into pyridoxal-5-phosphate by activating the pyridoxal 5-phosphate-kinase enzyme, in contrast to other phospholipids of different origins. The possible role of brain cortex phospholipids in regulating an enzyme contained in the soluble fraction is discussed. Moreover, it is suggested that the specific effect of the phospholipids from various sources is linked to their fatty acid composition and to be therefore dependent on the aliphatic chains.     

p82H identifies sequences at every human centromere

Summary A cloned alphoid sequence, p82H, hybridizes in situ to the centromere of every human chromosome. After washing under stringent conditions, no more than 8% of the grains are located on any specific chromosome. p82H thus differs from other centromeric sequences which are reported to be chromosome specific, because it detects sequences that are conserved among the chromosomes. Two experimental approaches show that the p82H sequences are closely associated with the centromere. First, p82H remains with the relocated centromeres in an inv(19) and an inv(6) chromosome. Second, p82H hybridizes at the centromere but not to the centromeric heterochromatin of chromosomes 1, 9 and 16 that have elongated 1qh, 9qh and 16qh regions produced by short growth in 5-azacytidine. The only noncentromeric site of hybridization is at the distal end of the 9qh region.     

Phytochrome-mediated responses of cells and protoplasts of green calli obtained from the leaves of a CAM plant

Green callus obtained from leaves of the CAM-inducible plant Kalanchoe blossfeldiana cv. Montezuma has previously been shown to perform C₃-type photosynthesis under 16-h days and to shift to crassulacean acid metabolism (CAM) under 9-h days. The utilization of photoperiodic regimes (i.e. night interruptions by 30 min red light) established that CAM induction in the callus was under the control of phytochrome, as shown by measurements of CAM criteria: phosphoenolpyruvate carboxylase activity and malic acid pools. Short-term responsiveness of the callus cells to phytochrome modulations by monochromatic radiations was also established by the rapid changes observed in the diameter of the callus-derived protoplasts. These results provide further evidence that whole plant correlations are not necessary for phytochrome operativity.Abbreviations CAM crassulacean acid metabolism - PAL phenylalanine ammonia lyase (EC 4.3.1.5) - PAR photosynthetically active radiations - PEPC phosphoenolpyruvate carboxylase (EC 4.1.1. 31) - Rubisco ribulose 1,5 bisphosphate carboxylase (EC 4.1.1.39)     

Chromosome-specific subsets of human alphoid DNA identified by a chromosome 2-derived clone

We have cloned an alphoid DNA fragment, pBS4D, from the DNA of a human-hamster hybrid cell line containing chromosome 2 as its only cytologically detectable human component. Under high stringency conditions, pBS4D hybridized in situ mostly to chromosome 2 and to a lesser extent to chromosomes 18 and 20. Restriction analysis using the DNA from selected somatic hybrid cell lines revealed that the genomic organization of this alphoid DNA differs on each of these three chromosomes.     

Holozygosity for sex-linked genes in males of the termiteIncisitermes schwarzi

The termite Incisitermes schwarzi has multiple sex chromosomes that have arisen by repeated translocations between autosomes and previously existing sex chromosomes. Two sex-linked allozyme loci--Acp-1 and Est-3--are holozygous, not hemizygous, in males (the heterogametic sex). Both loci show less than 1% crossing-over between X and Y chromosomes, and alleles of both are in marked disequilibrium with respect to X vs Y linkage. The two loci assort independently in female meiosis, indicating that they lie on different sex chromosomes. But they are tightly linked in male meiosis because of nonrandom assortment of the multiple X and Y chromosomes in males of this species. The findings of holozygosity and strong linkage disequilibrium suggest that differential selection in the two sexes at or near these loci may be responsible for the establishment of the translocations in this species. The existence of active Y-linked alleles also suggests that the translocations may have occurred recently.