Cell kinetic studies of chick brain cells in culture: an autoradiographic study with [3H]- and [14C]-thymidine

Labelling index, S-phase duration and cell-cycle time of proliferating brain cells from 6-day-old chick embryos in culture were investigated autoradiographically after labelling with [3H]- and/or [14C]-thymidine. The dissociated cells were cultured in the absence or in the presence of brain extract from 8-day-old chick embryos. Cultures contained essentially two cell types, which could be easily distinguished by the size of their nuclei: small nuclei identified as belonging to precursor cells of neurons and large nuclei corresponding to astroglial cells. The labelling index of astroglial cells (16.4%) was about 2 times higher than that of the neuronal cells (9.9%). Under the influence of brain extract the labelling index of neuroblasts was nearly doubled while that of the astroglial cells remained nearly unchanged. From double-labelling experiments with [3H]- and [14C]-thymidine, the same S-phase duration of about 7 hr was found for both cell types cultured with or without brain extract. A cell-cycle duration of 39 hr for neuronal and of 29 hr for astroglial cells was found. The cycle times remained constant under the influence of brain extract. From the measured data mentioned above, a growth fraction of 50% (neuroblasts) and 68% (astroglial cells) was calculated in control cultures without brain extract. After addition of brain extract, the growth fraction increased for both cell types (neuroblasts: 92%; astroglial cells: 80%). The results demonstrate that more cells proliferate in the presence of brain extract, but the durations of the S-phase and the cell cycle remain unchanged.     

Ecophysiological and species‐specific responses to seasonal variations in halophytic species of the chenopodiaceae in a Mediterranean salt marsh

Plants adopt different strategies in response to seasonal variations in their natural biotype. In this study, the responses of seasonal changes on the organic osmolyte contents, ionic homoeostasis, osmotic potential and succulence of native chenopods in a salt marsh of Egypt were studied. Five halophytic species of the chenopodiaceae were collected during the rainy and dry season: Atriplex halimus L., Halocnemum strobilaceum (Pall.), Salicornia fruticosa (L.), Suaeda pruinosa (Lange) and Arthrocnemum macrostachyum (Moric). Results showed that plants employ an osmoconformer strategy via species specificity of response to seasonal variations. The total ions accumulation in all species was higher during the rainy season compared with the dry season. In particular, during rainy season, A. halimus resulted the higher accumulator species, followed by H. strobilaceum. Conversely, S. fruticosa, S. pruinosa and A. macrostachyum were defined as low‐accumulator species. In each studied species, the effective salinity, biological absorption coefficient as well as leaf succulence were higher during the rainy season. In conclusion, the present study have pointed out the importance that should be accorded with the development of research directed towards agronomic aspects for the uses of halophytic chenopodiaceae in salt marsh. Further, data obtained indicate that the ions accumulator species should be considered for use in phytoremediation of degraded saline lands.     

Monitoring treatment response of childhood acute lymphocytic leukemia with certain molecular and biochemical markers

Apoptosis is the primary mechanism through which most chemotherapeutic agents induce tumor cell death. The purpose of this study was to monitor the expression of pro- and anti-apoptotic proteins CD(95) , Bcl-2, as well as copper and zinc levels in the peripheral blood of children with acute lymphocytic leukemia (ALL) prior to and 6 months after the beginning of chemotherapy. Blood parameters and bone marrow blast count were also assessed. Twenty of 26 patients who received treatment showed amelioration in apoptotic response, which is reflected in the elevation of CD(95) , whereas Bcl-2 protein was significantly lowered. In these patients, the elevated serum copper level was not significantly affected whereas the low serum zinc level was significantly raised. Improvement in blood parameters and bone marrow blast count were also achieved. Taken together, the data suggested that assessment of apoptosis signaling molecules might have a predictive impact on treatment outcome.     

Anatomical significance of the hygrochastic movement in Anastatica hierochuntica

BACKGROUND AND AIMS: Unlike the dispersal mechanisms of many desert plants, the whole dead skeleton of Anastatica hierochuntica is involved in seed dispersal and preservation. This process depends on the hygrochastic nature of the lignified conducting tissue that bends when dry and straightens under wet conditions. An anatomical interpretation of this mechanical movement was investigated. METHODS: An anatomical study of the stem was conducted on the juvenile plants raised under different water treatments and on the branch-orders of adult A. hierochuntica size-classes. KEY RESULTS: In the juvenile stem of A. hierochuntica, the area of cortex, conducting tissue and pith increased with water availability. However, the hydraulic conductance decreased, resulting in a better withdrawal of water in water-stressed plants. The anatomical investigation of the hygrochastic mechanism revealed an asymmetric distribution of the cortical tissues, with the conducting tissues of the stem of juvenile and adult plants being larger in the lower side. The hydraulic conductance was better in the basal and middle branch-orders than the terminal ones, permitting better conductance of water to the subsequent branch-orders. CONCLUSIONS: The lignified conducting tissue of the whole stem, having a hygrochastic nature, controls the movement of the branches. The greater amount of conducting tissue associated with a higher density of wide xylem vessels was observed in the lower side of the stem as compared with the upper side. Consequently, the conducting tissue in the lower side of the stem was suggested to be more effective in the opening process of the curled dry branches through better and more rapid conductance of water. Alternatively, due to the few narrow xylem vessels in the upper side of the stem, it was likely that the conducting tissue in the upper side is more effective in the closing process by providing more rapid drying. The mechanical rise of water and the related hygrochastic efficiency were maximized in the basal and middle branch-orders that are mostly involved in the mechanical movement.     

Urethrographic examinations: Patient and staff exposures and associated radiobiological risks

Medical exposure of the general population due to radiological investigations is the foremost source of all artificial ionising radiation. Here, we focus on a particular diagnostic radiological procedure, as only limited data are published with regard to radiation measurements during urethrograpic imaging. Specifically, this work seeks to estimate patient and occupational effective doses during urethrographic procedures at three radiology hospitals. Both staff and patient X-ray exposure levels were calculated in terms of entrance surface air kerma (ESAK), obtained by means of lithium fluoride thermoluminescent dosimeters (TLD-100(LiF:Mg:Cu.P)) for 243 urethrographic examinations. Patient radiation effective doses per procedure were estimated using conversion factors obtained from the use of Public Health England computer software. In units of mGy, the median and range of ESAK per examination were found to be 10.8 (3.6–26.2), 7.0 (0.2–32.3), and 24.3 (9.0–32.0) in Hospitals A, B, and C, respectively. The overall mean and range of staff doses (in µGy) were found to be 310 (4.0–1750) per procedure. With the exception of hospital C, the present evaluations of radiation dose have been found to be similar to those of previously published research. The wide range of patient and staff doses illustrate the need for radiation dose optimisation.     

Examining requirement for formation of functional Presenilin proteins and their processing events in vivo

Presenilin (Psn) is a multipass transmembrane protein that functions as the catalytic subunit of γ‐secretase for mediating intramembrane cleavage of type 1 transmembrane proteins. Normally active Psn is in the form of a heterodimer composed by its N‐terminal and C‐terminal fragments that are generated from a Presenilinase‐mediated endoproteolytic cleavage within its large cytosolic loop during assembly of the protease complex. Using the Psn forms that either bypass or disable Presenilinase‐mediated endoproteolysis, and a Psn form that has most of the large cytosolic loop deleted, we have established an in vivo system to enable investigations of Psn functional domains in Drosophila. We show that the Presenilinase‐mediated endoproteolytic event is not essential for producing Psn activity during animal development, and is regulated by integrity of the large cytosolic loop of Psn in Drosophila. The Psn transgenic flies described here could be applied to a broad range of studies on Psn functioning and its related γ‐secretase activity at any developmental stage. genesis 47:161–168, 2009. © 2009 Wiley‐Liss, Inc.     

The small GTPase BcCdc42 affects nuclear division, germination and virulence of the gray mold fungus Botrytis cinerea

The small GTPase Cdc42 plays a central role in various processes in eukaryotic cells including growth, differentiation and cytoskeleton organization. Whereas it is essential in the yeast Saccharomyces cerevisiae, its role in filamentous fungi differs, due to the complementing, partly overlapping function of Rac. We analyzed the role of the Cdc42 homologue in the necrotrophic, broad host range pathogen Botrytis cinerea. Deletion mutants of bccdc42 showed various growth abnormalities; the mutants had reduced growth rate and hyphal branching, they produced fewer conidia, which were enlarged and misshapen and had germination defects. Additionally, the mutants were impaired in sclerotia development. Cytological studies indicate that at least part of this phenotype could be attributed to disturbed control of nuclear division: conidia and hyphae of the mutant showed twofold higher nucleus/cytoplasm ratio compared to wild type cells. Apart from these effects on vegetative growth and differentiation, Δbccdc42 strains were attenuated in penetration and colonization of host tissue, confirming that BcCdc42 – though being not essential like in yeast – is involved in important developmental processes in B. cinerea.     

Generation of a large-scale genomic resource for functional and comparative genomics in Liriodendron tulipifera L.

Liriodendron tulipifera L., a member of Magnoliaceae in the order Magnoliales, has been used extensively as a reference species in studies on plant evolution. However, genomic resources for this tree species are limited. We constructed cDNA libraries from ten different types of tissues: premeiotic flower buds, postmeiotic flower buds, open flowers, developing fruit, terminal buds, leaves, cambium, xylem, roots, and seedlings. EST sequences were generated either by 454 GS FLX or Sanger methods. Assembly of almost 2.4 million sequencing reads from all libraries resulted in 137,923 unigenes (132,905 contigs and 4,599 singletons). About 50% of the unigenes had significant matches to publically available plant protein sequences, representing a wide variety of putative functions. Approximately 30,000 simple sequence repeats were identified. More than 97% of the cell wall formation genes in the Cell Wall Navigator and the MAIZEWALL databases are represented. The cinnamyl alcohol dehydrogenase (CAD) homologs identified in the L. tulipifera EST dataset showed different expression levels in the ten tissue types included in this study. In particular, the LtuCAD1 was found to partially recover the stiffness of the floral stems in the Arabidopsis thaliana CAD4 and CAD5 double mutant plants, of the LtuCAD1 in lignin biosynthesis. L. tulipifera genes have greater sequence similarity to homologs from other woody angiosperm species than to non-woody model plants. This large-scale genomic resour"HistryDatesce will be instrumental for gene discovery, cDNA microarray production, and marker-assisted breeding in L. tulipifera, and strengthen this species' role in comparative studies.