Identification to the species level of <Emphasis Type="Italic">Lactobacillus</Emphasis> isolated in probiotic prospecting studies of human,animal or food origin by 16S-23S rRNA restriction profiling

Background  

The accurate identification of Lactobacillus and other co-isolated bacteria during microbial ecological studies of ecosystems such as the human or animal intestinal tracts and food products is a hard task by phenotypic methods requiring additional tests such as protein and/or lipids profiling.     

Morphological differences among egg nests and adult individuals of Cicadatra persica (Hemiptera,Cicadidae), distributed in Erneh,Syria

The aim of this study is determining the different patterns of egg nests and the morphological differences between the specimens of Cicadatra persica Kirkalidy, 1909 (Hemiptera: Cicadidae) distributed in fruit orchards in Erneh located on AL-Sheikh mountain south west of Syria. The appearance of 80 egg nests was studied, and the results showed that there were two basic patterns of egg nests laid by Cicadatra persica, 90% of the egg nests were of the first pattern (consists of several adjacent slits), while 10% of them were of the second pattern (consists of several divergent slits). A random sample consisting of 300 specimens (150 males and 150 females) were also studied concentrating on the differences in the color of the supra-antennal plate and in the number of spurs on the tibia of the hind legs. The results showed that there were two basic patterns of individuals based on the differences in the color of supra-antennal plate. The first pattern (individuals with yellow supra-antennal plates), constituted more than 90%, and the second one (individuals with black supra-antennal plates) constituted less than 10%. The results also showed that there were 27 different patterns based on the number of spurs on the tibia of the hind legs. One of them was a common pattern (2, 3) whose individuals have 2 spurs on the upper side of the tibia of the hind legs and 3 spurs on the lateral side of the tibia of the hind legs. The total percent of this common pattern was 76%. The other 26 patterns were different from each other, and the total percent of all these different patterns was 24%.     

Improving the yellow pigment content of bread wheat flour by selecting the three homoeologous copies of Psy1

The yellow pigment content (YPC) of endosperm affects the quality and nutritional value of wheat grain products. Major quantitative trait loci (QTL) for endosperm YPC have been repeatedly mapped on chromosomes 7A and 7B in durum and bread wheats. The genes coding for phytoene synthase (PSY1), which is involved in the biosynthesis of carotenoids, generally co-segregate with these QTL, indicating their role in determining YPC. Here, to study the genetic factors underlying endosperm YPC in bread wheat, the sequence polymorphism of the homoeologous A, B and D copies of genes coding for PSY1, Psy-A1, Psy-B1, and Psy-D1, was studied in a worldwide core collection, which was also phenotyped for flour YPC. Seven novel alleles of Psy-A1 and two novel alleles of Psy-B1 were detected, which confirms the high level of polymorphism of these genes. Two major QTL with respective candidate genes Psy-A1 and Psy-B1 were identified in the distal region of chromosomes 7A and 7B using progeny of a cross between Apache and Ornicar, high and low YPC cultivars, respectively. Association mapping confirms the role of these genes in YPC and shows that the D copy also significantly influences this trait. These results indicate that breeders need to consider all three Psy1 copies when seeking to improve the YPC of wheat endosperm.     

Collagenase predigestion on paraffin sections enhances collagen immunohistochemical detection without distorting tissue morphology

Reliable immunohistochemical detection of collagen in formalin fixed, paraffin embedded tissues requires protease digestion. While these pan-proteases (pepsin, trypsin, protease K, etc.) enhance collagen detection, they also digest many other tissue proteins and produce poor cellular morphology and unrecognizable cellular structures. Balancing the conditions (protease type, concentration, incubation time and temperature) to digest some, but not all, proteins in a tissue section while optimizing collagen detection requires one to compromise improved collagen immunolabeling with adequate cellular morphology. Furthermore, optimal conditions for digesting tissue proteins to enhance collagen detection vary among tissue types and their fixation. Although brain is not typically subject to these deleterious consequences, structures such as epithelium, spermatids, stroma etc. and other tissues with complicated histology are profoundly affected. To resolve this technical dilemma, we discovered a novel use for collagenase to enhance collagen immunodetection without affecting the noncollagen proteins, thereby preserving tissue morphology. Collagenase, which is typically used in vitro for disassociation of cells, has never been used reliably on formalin fixed, paraffin embedded tissue sections. This new use of collagenase for immunohistochemistry promotes increased collagen immunolabeling, is easy to use, is versatile, and allows preservation of tissue structure that provides maximal and accurate histological information.     

Inheritance of the number and thickness of cell layers in barley aleurone tissue (<Emphasis Type="Italic">Hordeum vulgare </Emphasis>L.): an approach using F2–F3 progeny

The aleurone tissue of cereal grains, nutritionally rich in minerals and vitamins, is an important target for the improvement of cereals. Inheritance of the thickness and the number of cell layers in barley aleurone was studied on the F2–F3 progeny of an Erhard Frederichen × Criolla Negra cross in which the parental lines have three or two aleurone layers, respectively. F3 grain was sampled from each F2 plant and 96.8% of the entire F3 grain population was classified as being either the 2- or 3-layer type. Using microsatellite, single nucleotide polymorphism (SNP) and morphological markers on 190 F2 plants, a linkage map was built. Three quantitative trait loci (QTLs) affecting aleurone traits were revealed on chromosome 5H (max. LOD = 5.83) and chromosome 7H (max. LOD = 4.45) by interval mapping, and on chromosome 2H by marker analysis with an unmapped marker. These QTLs were consistent with genetic sub-models involving either 2-cell type dominance for 7H and 2H, or putative partial dominance for 5H where 2-cell-layer dominance and additivity gave similar LODs. The number of aleurone cell layers and aleurone thickness were strongly correlated and QTL results for these traits were alike. An SNP marker of sal1, an orthologue of the maize multilayer aleurone gene was mapped to the 7HL chromosome arm. However, the 7H QTL did not co-locate with the barley sal1 SNP, suggesting that an additional gene is involved in determining aleurone traits. These new mapping data allow comparisons to be made with related studies.     

Investigation of apoptosis in cultured cells infected with equine herpesvirus 1

Many viruses alter different stages of apoptosis of infected cells as a strategy for successful infection. Few studies have addressed mechanisms of equine herpesvirus 1 (EHV-1) strain-induced cell death. We investigated the effect of an abortigenic strain (AR8 strain) on heterologous Madin–Darby bovine kidney cells and homologous equine dermis (ED) cells cell lines. We compared morphologic and biochemical features of early and late apoptosis at different postinfection times. We investigated translocation of phosphatidylserine to the cell surface, nuclear fragmentation and changes in the cytoskeleton using flow cytometry and annexin V/propidium iodide staining, DNA laddering, terminal deoxynucleotidyl transferase UTP nick-end labeling assay and immunofluorescence staining of cytokeratin 18 cleavage. AR8 EVH-1 strain interfered with apoptosis in both cell lines, particularly during the middle stage of the replication cycle; this was more evident in ED cells. Although this antiapoptotic effect has been reported for other alpha herpesviruses, our findings may help elucidate how EHV-1 improves its infectivity during its cycle.     

Stent implantation through a self-expanding stent

Jailing of a side-branch is a known complication of stent implantation, and makes access to the side-branch difficult, especially if the stent is of the self-expanding type. Although plain balloon angioplasty is feasible for the jailed side-branches, the use of newer devices (a stent, Rotablation or atherectomy) has not been described. We describe a novel way of treating a side-branch jailed by a self-expanding stent by using stent implantation through the strut of a self-expanding stent.