Collection of Epithelial Cells from Rodent Mammary Gland Via Laser Capture Microdissection Yielding High-Quality RNA Suitable for Microarray Analysis

Laser capture microdissection (LCM) enables collection of cell populations highly enriched for specific cell types that have the potential of yielding critical information about physiological and pathophysiological processes. One use of cells collected by LCM is for gene expression profiling. Samples intended for transcript analyses should be of the highest quality possible. RNA degradation is an ever-present concern in molecular biological assays, and LCM is no exception. This paper identifies issues related to preparation, collection, and processing in a lipid-rich tissue, rodent mammary gland, in which the epithelial to stromal cell ratio is low and the stromal component is primarily adipocytes, a situation that presents numerous technical challenges for high-quality RNA isolation. Our goal was to improve the procedure so that a greater probe set present call rate would be obtained when isolated RNA was evaluated using Affymetrix microarrays. The results showed that the quality of RNA isolated from epithelial cells of both mammary gland and mammary adenocarcinomas was high with a probe set present call rate of 65% and a high signal-to-noise ratio

Is Evolution of Blind Mole Rats Determined by Climate Oscillations?

The concept of climate variability facilitating adaptive radiation supported by the ‘‘Court Jester’’ hypothesis is disputed by the ‘‘Red Queen’’ one, but the prevalence of one or the other might be scale-dependent. We report on a detailed, comprehensive phylo-geographic study on the ,4 kb mtDNA sequence in underground blind mole rats of the family Spalacidae (or subfamily Spalacinae) from the East Mediterranean steppes. Our study aimed at testing the presence of periodicities in branching patterns on a constructed phylogenetic tree and at searching for congruence between branching events, tectonic history and paleoclimates. In contrast to the strong support for the majority of the branching events on the tree, the absence of support in a few instances indicates that network-like evolution could exist in spalacids. In our tree, robust support was given, in concordance with paleontological data, for the separation of spalacids from muroid rodents during the first half of the Miocene when open, grass-dominated habitats were established. Marine barriers formed between Anatolia and the Balkans could have facilitated the separation of the lineage ‘‘Spalax’’ from the lineage ‘‘Nannospalax’’ and of the clade ‘‘leucodon’’ from the clade ‘‘xanthodon’’. The separation of the clade ‘‘ehrenbergi’’ occurred during the late stages of the tectonically induced uplift of the Anatolian high plateaus and mountains, whereas the separation of the clade ‘‘vasvarii’’ took place when the rapidly uplifting Taurus mountain range prevented the Mediterranean rainfalls from reaching the Central Anatolian Plateau. The separation of Spalax antiquus and S. graecus occurred when the southeastern Carpathians were uplifted. Despite the role played by tectonic events, branching events that show periodicity corresponding to 400-kyr and 100-kyr eccentricity bands illuminate the important role of orbital fluctuations on adaptive radiation in spalacids. At the given scale, our results supports the ‘‘Court Jester’’ hypothesis over the ‘‘Red Queen’’ one

Molecular characterization of MHC class IIB genes of sympatric Neotropical cichlids

Ministerio de Economía y Competitividad del Gobierno de España, Programa de Formación de Personal Investigador FPI BES-2011-047645 to MJH, Programa Estatal de Fomento de la Investigación Científica y Técnica de Excelencia Proyecto CGL 2010-16103 to MB. This project was further enabled through two German Science Foundation grants to CE (DFG, EI841/4-1 and EI841/6-1) both part of the SPP 1399 priority programme on “host-parasite interactions”

Morphological examination of Hungarian apricot rootstock varieties

Apricot seedlings are one of the most widely used rootstocks in apricot growing of Hungary. The National List of Hungary contains 7 apricot seedling rootstocks varieties. The production of apricot graft on seedling rootstock takes 3 years from the seed production to the complete graft and requires more space. It is important to provide the trueness to variety name during this process. In the genus Prunoideae not only the plant but also the stone should have typical characteristics. It seems to be useful to examine the morphology of stone. The observed varieties were 'Tengeribarack C.1300', 'Tengeribarack C.1301', 'Tengeribarack C.1650', 'Tengeribarack' C.1652', 'Tengeribarack C.145', 'Tengeribarack C.1426' and 'Tengeribarack C.2546`.1t seems to be the most suitable characteristic to make a distinction between these varieties by stone, the ratio height of stone to lateral width of stone, the ratio of height of stone to ventral width of stone, the ratio of lateral width of stone to ventral width of stone and the ratio of ventral zone to ventral width. From not measured characteristic seems to be most utilizable to make distinction: the fusion of dorsal groove margins, the shape of apex, the presence of a mucro of the apex, the shape of stalk end in lateral view and the texture of lateral surfaces

Morphological examination of Hungarian apricot rootstock varieties

Apricot seedlings are one of the most widely used rootstocks in apricot growing of Hungary. The National List of Hungary contains 7 apricot seedling rootstocks varieties. The production of apricot graft on seedling rootstock takes 3 years from the seed production to the complete graft and requires more space. It is important to provide the trueness to variety name during this process. In the genus Prunoideae not only the plant but also the stone should have typical characteristics. It seems to be useful to examine the morphology of stone. The observed varieties were 'Tengeribarack C.1300', 'Tengeribarack C.1301', 'Tengeribarack C.1650', 'Tengeribarack' C.1652', 'Tengeribarack C.145', 'Tengeribarack C.1426' and 'Tengeribarack C.2546`.1t seems to be the most suitable characteristic to make a distinction between these varieties by stone, the ratio height of stone to lateral width of stone, the ratio of height of stone to ventral width of stone, the ratio of lateral width of stone to ventral width of stone and the ratio of ventral zone to ventral width. From not measured characteristic seems to be most utilizable to make distinction: the fusion of dorsal groove margins, the shape of apex, the presence of a mucro of the apex, the shape of stalk end in lateral view and the texture of lateral surfaces

MHC class I gene organization in > 1.5-Mb YAC contigs from the H2-M region.

Sixteen yeast artificial chromosome (YAC) clones have been mapped to the H2-M region at the distal end of the mouse major histocompatibility complex (MHC) on chromosome 17. Analysis of the YACs with single- and multicopy probes yielded a proximal contig spanning a minimum of 800 kb and a distal contig of 700 kb. A probe for the conserved fourth exon of MHC class I genes detected 19 restriction fragments, including 6 of the 8 previously characterized H2-M class I genes, in the proximal contig. This contig spans the gap from the M to the T region and includes the T1 gene. By contrast, only two class I genes, M2 and M3, were found in the distal contig. These two genes, which are both expressed, may mark the end of the MHC. The order among nine class I genes and seven other markers was determined in the cloned DNA from the centromere as T1, Tu32A, (M1-M7-M8), Tu32B, B30, M6, M4, M5, Mog, Tu42A parallel M2, Leh525, M3, Tu42B, where the orientation with respect to the centromere is unknown for M1-M7-M8