Irregular Cell Shape Research Articles

Dynamic crushing behavior of honeycomb structures with irregular cell shapes and non-uniform cell wall thickness

Dynamic crushing responses of honeycomb structures having irregular cell shapes and non-uniform cell wall thickness are studied using the Voronoi tessellation technique and the finite element (FE) method. FE models are constructed for such honeycomb structures based on Voronoi diagrams with different degrees of cell shape irregularity and cell wall thickness non-uniformity. The plateau stress, the densification strain energy and the initiation strain are determined using the FE models. Simulation results reveal that the “X” and “V” shaped deformation modes evident in a perfectly ordered honeycomb at low or moderate impact velocities are disrupted as cell shapes become irregular and/or cell wall thickness gets non-uniform. The “I” shaped deformation mode is clearly seen in all honeycomb structures at high impact velocities. Both the plateau stress and the densification strain energy are found to decrease as the degree of cell shape irregularity or the degree of cell wall thickness non-uniformity increases, with the weakening effect induced by the presence of non-uniform cell wall thickness being more significant. When the two types of imperfections co-exist in a honeycomb structure, the interaction between them is seen to exhibit a complicated pattern and to have a nonlinear effect on both the plateau stress and the densification strain energy. It is also found that stress waves propagate faster in a honeycomb structure having irregular cell shapes and slower in a honeycomb structure having non-uniform cell wall thickness than in a perfectly ordered honeycomb. Finally, the strain hardening of the cell wall material is seen to have a strengthening effect on the plateau stress, which is more significant for perfectly ordered honeycombs than for imperfect honeycomb structures.

Characterization of Hematopoietic Progenitor Cells Derived from Human Embryonic Stem Cells That Differentiate into Natural Killer Cells Capable of In Vivo Anti-Tumor Activity.

Derivation of multiple blood cell lineages from human embryonic stem cells (hESCs) clearly establishes these cells as an important model system to better characterize human hematopoietic ontogeny. Previously we have demonstrated development of myeloid, erythroid and lymphoid cells, as well as endothelial cells, from hESCs. However, the progenitor cells that give rise to these more mature cells remain poorly characterized. Here we use combined phenotypic and genetic analysis to discriminate between distinct progenitor cell populations. We also demonstrate that natural killer (NK) cells derived from these hESC-derived progenitors are capable of killing diverse tumors both in vitro and in vivo. By detailed flow cytometric analysis of differentiated hESCs over a defined time course, we identify development of two waves of CD34+ cells. The first wave consists of CD34brightCD31+Flk1+ cells, whereas the second wave consists of CD34dimCD45+ cells. Novel combined morphologic and phenotypic analysis by image scanning flow cytometry demonstrates these phenotypically different cell populations also are morphologically distinct. CD34brightCD31+ cells derived from hESCs appear to have a heterogeneous cell morphology, with an irregular cell shape, higher mean cell size and complex cytoplasmic organization. In contrast, CD34dimCD45+ cells are a more homogeneous cell population with a uniform spherical morphology and smaller cell size. Purified CD34brightCD31+Flk1+ cells express transcription factors associated with both the hematopoietic and endothelial lineages and can differentiate into both these lineages in vitro. In contrast, CD34dimCD45+ cells display a transcription profile suggestive of hematopoietic commitment and are significantly enriched for hematopoietic progenitors. Taken together, these results suggest that the CD34brightCD31+Flk1+ and CD34dimCD45+ cell populations are distinct cell lineages that represent early hemato-endothelial and hematopoietic precursors, respectively. Next, we better characterized the lymphoid developmental potential of the CD34dimCD45+ cells by the ability of these cells to differentiate into NK cells. Using a two-step differentiation process we find that hESC-derived NK cells express the wide repertoire of activating and inhibitory receptors similar to NK cells derived from other sources. We now demonstrate that these hESC-derived NK cells acquire cytolytic activity against breast cancer and glioma cell lines, as well as leukemia and lymphoma cells, in vitro. Finally, we have begun to test the in vivo efficacy of hESC-derived NK cells against established tumors. Here, luciferase (luc)-expressing K562 cells are inoculated into NOD/SCID mice, and several days later 2x106 NK cells derived from hESCs or from umbilical cord blood (UCB) cells are injected iv. The luc+ K562 cells allows serial bioluminescent imaging to follow growth of the tumor cells non-invasively over a prolonged time course. These studies demonstrate a beneficial effect of hESC-derived NK cells on tumor growth and metastasis, comparable to the effect of UCB-derived NK cells. Taken together, these studies characterize the earliest hematopoietic progenitors as they develop from hESCs during in vitro differentiation, providing a starting point to evaluate the effect of endogenous and exogenous factors on differentiation of distinct human hematopoietic lineages. This will translate into even more efficient derivation of NK cells from hESCs with a potential use for cancer immunotherapy.

Rice NTRC is a high-efficiency redox system for chloroplast protection against oxidative damage.

One of the mechanisms plants have developed for chloroplast protection against oxidative damage involves a 2-Cys peroxiredoxin, which has been proposed to be reduced by ferredoxin and plastid thioredoxins, Trx x and CDSP32, the FTR/Trx pathway. We show that rice (Oryza sativa) chloroplast NADPH THIOREDOXIN REDUCTASE (NTRC), with a thioredoxin domain, uses NADPH to reduce the chloroplast 2-Cys peroxiredoxin BAS1, which then reduces hydrogen peroxide. The presence of both NTR and Trx-like domains in a single polypeptide is absolutely required for the high catalytic efficiency of NTRC. An Arabidopsis thaliana knockout mutant for NTRC shows irregular mesophyll cell shape, abnormal chloroplast structure, and unbalanced BAS1 redox state, resulting in impaired photosynthesis rate under low light. Constitutive expression of wild-type NTRC in mutant transgenic lines rescued this phenotype. Moreover, prolonged darkness followed by light/dark incubation produced an increase in hydrogen peroxide and lipid peroxidation in leaves and accelerated senescence of NTRC-deficient plants. We propose that NTRC constitutes an alternative system for chloroplast protection against oxidative damage, using NADPH as the source of reducing power. Since no light-driven reduced ferredoxin is produced at night, the NTRC-BAS1 pathway may be a key detoxification system during darkness, with NADPH produced by the oxidative pentose phosphate pathway as the source of reducing power.

Ultrastructural examination of the host inflammatory response within gills of netpen reared chinook salmon ( Oncorhynchus tshawytscha) with Microsporidial Gill Disease

The sequence of host changes following the rupture of spore-laden xenomas of the microsporidian Loma salmonae during Microsporidial Gill Disease of Salmon was deduced from ultrastructural examination of the gills of naturally infected, moribund, chinook salmon from a commercial aquaculture site. The gills contained many stages of parasite development suggesting fish were chronically exposed to the parasite. Intact xenomas were generally found beneath the endothelium in arteries and arterioles and were encapsulated by a layer of collagen containing fibroblasts sometimes joined by desmosomes. Xenoma dissolution was characterized by neutrophil infiltration and loss of the xenoma plasma membrane and encapsulation. The inflammatory responses associated with ruptured xenomas ranged from acute lesions, denoted by a marked neutrophil infiltration and vascular thrombosis, to chronic lesions with a macrophage-rich infiltrate variously accompanied by neovascularization and vascular remodelling. Dendritic-like cells and plasma cells were characteristic throughout. Basement membrane damage of the primary filament epithelium and subsequent transepithelial expulsion of spores were associated with severe inflammation. An unusual previously undescribed multifocal change, in which epithelial cells invaded deeply beyond the normal boundaries of the basement membrane, affected areas of gill filament epithelium with basement membrane damage. Some neutrophils that contained L. salmonae spores, or spore polar tube, displayed morphological changes that included irregular cell shape, cytoplasmic darkening associated with an abundance of free ribosomes, lysis of neighbouring cells, and type II nuclear clefts. Fusion of apparently intact neutrophils occurred in other areas of the lesion, where close contacts between neighbouring cells were established and in some areas plasma membrane fusion occurred. Closely associated neutrophils with intact plasma membranes were observed to contain type II nuclear clefts, abundant granules and rough endoplasmic reticulum. Other neutrophils in the lesion displayed type I nuclear pockets, which is suspected to be an early stage of apoptosis.

Numerical Determination of Transmembrane Voltage Induced on Irregularly Shaped Cells

The paper presents an approach that reduces several difficulties related to the determination of induced transmembrane voltage (ITV) on irregularly shaped cells. We first describe a method for constructing realistic models of irregularly shaped cells based on microscopic imaging. This provides a possibility to determine the ITV on the same cells on which an experiment is carried out, and can be of considerable importance in understanding and interpretation of the data. We also show how the finite-thickness, nonzero-conductivity membrane can be replaced by a boundary condition in which a specific surface conductivity is assigned to the interface between the cell interior (the cytoplasm) and the exterior. We verify the results obtained using this method by a comparison with the analytical solution for an isolated spherical cell and a tilted oblate spheroidal cell, obtaining a very good agreement in both cases. In addition, we compare the ITV computed for a model of two irregularly shaped CHO cells with the ITV measured on the same two cells by means of a potentiometric fluorescent dye, and also with the ITV computed for a simplified model of these two cells.

Cytokines and Vascular Permeability: An In Vitro Study on Human Endothelial Cells in Relation to Tumor Necrosis Factor-α-Primed Peripheral Blood Mononuclear Cells

Tumor response is strongly enhanced by addition of tumor necrosis factor (TNF)-alpha to chemotherapy in local-regional perfusion. TNF primarily targets the endothelial lining of the tumor-associated vasculature, thereby improving permeability of the vascular bed. This augments uptake of the coadministered chemotherapeutic drug in the tumor. In vitro, however the high dose of TNF did not directly affect endothelial cells, indicating that other factors, most likely TNF-induced, are involved in the antivascular activities observed in vivo. This is supported by in vivo studies in our laboratory in which depletion of leukocytes resulted in loss of the antivascular activity of TNF. The present study examined the role of peripheral blood mononuclear cells (PBMCs) on endothelial cells by exposing them to TNF, interferon (IFN)-gamma, and PBMCs. We observed morphological changes of the endothelial cells when exposed to TNF in combination with IFN. Endothelial cells became elongated. and gaps between the cells formed. Addition of PBMCs enhanced these alterations. The endothelial layer became disrupted with highly irregular-shaped cells displaying large gap formations. PBMCs also contributed to an increased permeability of the endothelial layer without augmenting apoptosis. Replacing PBMC by interleukin (IL)-1beta produced similar effect with regard to inhibition of cell growth, morphological changes, and induction of apoptosis. Blocking IL-1beta with a neutralizing antibody diminished the effects inflicted of PBMCs. These observations indicate that endogenously produced IL-1beta by primed PBMCs plays an important role in the antivascular effect of TNF.

Pulmonary Intravascular Hemangiosarcoma in a Cat

A 9-year-old, castrated male Japanese domestic cat presented with a complaint of exertional dyspnea. Based on the radiographic findings, presumptive diagnosis of aspiration pneumonia or primary diffuse pulmonary neoplasia in the right middle lobe was made. Histologically, the pulmonary lesion was characterized by diffuse thickening of alveolar wall with the proliferation of apparently atypical irregular-shaped cells. Immunohistochemical staining using anti-human factor VIII-related antigen antiserum showed positive reaction in the cytoplasm of the atypical cells. According to the findings, the lesion was diagnosed as pulmonary intravascular hemangiosarcoma.

Effects of cell shape and strut cross-sectional area variations on the elastic properties of three-dimensional open-cell foams

The Voronoi tessellation technique and the finite element (FE) method are utilized to investigate the microstructure–property relations of three-dimensional (3-D) cellular solids (foams) that have irregular cell shapes and non-uniform strut cross-sectional areas (SCSAs). Perturbations are introduced to a regular packing of seeds to generate a spatially periodic Voronoi diagram with different degrees of cell shape irregularity (amplitude a), and to the constant SCSA to generate a uniform distribution of SCSAs with different degrees of SCSA non-uniformity (amplitude b). Twenty FE models are constructed, based on the Voronoi diagrams for twenty foam samples (specimens) having the same pair of a and b, to obtain the mean values and standard deviations of the elastic properties. Spatially periodic boundary conditions are applied to each specimen. The simulation results indicate that for low-density imperfect foams, the elastic moduli increase as cell shapes become more irregular, but decrease as SCSAs get less uniform. When the relative density ( R) increases, the elastic moduli of imperfect foams increase substantially, while the Poisson's ratios decrease moderately. The effect of the interaction between the two types of imperfections on foam elastic properties appears to be weak. In addition, it is found that the strut cross-sectional shape has a significant effect on the foam properties. Also, the elastic response of foams with the cell shape and SCSA imperfections appears to be isotropic regardless of changes in a, b and R and the strut cross-sectional shape.

Oncogenic K-Ras down-regulates Rac1 and RhoA activity and enhances migration and invasion of pancreatic carcinoma cells through activation of p38

Activating mutations in the K- ras gene are genetic alterations frequently found in human carcinomas, particularly in pancreatic adenocarcinomas. Mutation of the K- ras gene is thought to be an early and important event in pancreatic tumor initiation, but the precise role of the mutant K-Ras proteins in neoplastic progression is still unknown. In the present study, we have characterized the influence of oncogenic K-Ras on the phenotype and on the signal transduction of epitheloid PANC-1 pancreatic carcinoma cells by generating PANC-1 cell clones, which stably express EGFP(enhanced green fluorescent protein)-K-Ras (V12). EGFP-K-Ras (V12)-expressing cells exhibited a more fibroblastoid cellular phenotype with irregular cell shape and disorganized cytokeratin filaments. Moreover, these cells showed a marked enhancement of their migratory and invasive properties. Stable expression of EGFP-K-Ras (V12) down-regulated the activity of Rac1 and RhoA, resulting in reduced subcortical actin filaments and stress fibers, which might contribute to the epithelial dedifferentiation. Characterization of the activity of mitogen-activated protein kinases revealed that EGFP-K-Ras (V12) enhanced the activity of p38, but did not affect the activities of the Raf/MEK/ERK cascade and JNK. While inhibition of either MEK or JNK activity had no effect on EGFP-K-Ras (V12)-induced migration, inhibition of p38 activity markedly reduced EGFP-K-Ras (V12)-induced migration. Collectively, the results suggest that oncogenic K-Ras enhances the malignant phenotype and identify the mitogen-activated protein kinase p38 as a target to inhibit oncogenic K-Ras-induced pancreatic tumor cell migration.

INFLUENCE OF OZONATED WATER ON THE STRUCTURE AND SOME QUALITY PARAMETERS OF WHOLE STRAWBERRIES IN MODIFIED ATMOSPHERE PACKAGING (MAP)

Strawberry fruits (Fragaria x ananassa Duch. 'Darselect') were submerged in ozonated water (1.66 ppm) for 5 minutes, packed in air and in modified atmosphere (65% N 2 O, 25% Ar, 5% O 2 , 5% CO 2 ) and stored at 4°C for 20 days. SEM analyses of the surface cells of the fresh product before and after treatment were performed. Titratable acidity, color and texture modifications during storage were assessed. Color evaluation of surface of the fruit showed a rapid increase of red index (a*) and a decrease of luminosity values (L*) during storage, in particular for samples pretreated and packed in air. The results of color determinations, performed with image analysis on strawberry sepals, indicated a preservative effect of the ozonated water pretreatment. SEM images of the walls cells close to the fruit surface of untreated and treated samples, showed some irregular shape of cells of pretreated strawberries, compared to those of untreated fruit. The dynamometric results, obtained with texture analyzer, did not show significant differences in the decrease of maximum firmness between samples. However, the energy required to penetrate the surface layer (5 mm in depth) of treated and untreated strawberry samples differed significantly, confirming a more pronounced superficial softening in fruits pretreated with ozonated water. Moreover, the samples not ozonated and packed in MA showed, in comparison to control sample (packed in air), a reduction in rate of softening and a preservative effect on color during the early period of storage. Ozonated water pretreatments seem to be not effective in prolonging the shelf-life of strawberries, in particular in terms of maintenance of surface layer firmness.

Cortical Actin Turnover during Cytokinesis Requires Myosin II

The involvement of myosin II in cytokinesis has been demonstrated with microinjection, genetic, and pharmacological approaches; however, the exact role of myosin II in cell division remains poorly understood. To address this question, we treated dividing normal rat kidney (NRK) cells with blebbistatin, a potent inhibitor of the nonmuscle myosin II ATPase. Blebbistatin caused a strong inhibition of cytokinesis but no detectable effect on the equatorial localization of actin or myosin. However, whereas these filaments dissociated from the equator in control cells during late cytokinesis, they persisted in blebbistatin-treated cells over an extended period of time. The accumulation of equatorial actin was caused by the inhibition of actin filament turnover, as suggested by a 2-fold increase in recovery half-time after fluorescence photobleaching. Local release of blebbistatin at the equator caused localized accumulation of equatorial actin and inhibition of cytokinesis, consistent with the function of myosin II along the furrow. However, treatment of the polar region also caused a high frequency of abnormal cytokinesis, suggesting that myosin II may play a second, global role. Our observations indicate that myosin II ATPase is not required for the assembly of equatorial cortex during cytokinesis but is essential for its subsequent turnover and remodeling.

Irregular nuclear shape of bone marrow plasma cells defines a multiple myeloma subgroup related to hypodiploidy and to short survival

Morphological changes of plasma cells (PC) are common in multiple myeloma (MM). Loss of round or oval nuclear shape has been related to cell malignancy in human, and we looked for the occurrence of such morphological change on PC from bone marrow (BM) smears in a retrospective series of 169 MM patients at diagnosis. Nuclear shape changes of PC differed according to the patients (notch, dumb-bell, folded or monocytoid appearance), even in the same patient; all subtypes were pooled and defined as PC with irregular nuclear shape (PCIN). A significant number of PCIN (≥5% of all BMPC) was found at diagnosis in 20.7%. Median survival was of 22 months for patients with ≥5% PCIN, and 41 months for others ( p = 0.0001). Significant relationship was observed with prognostic parameters related intrinsic malignancy of the tumour process but not with beta-2-microglobulin (b2m). A clear-cut relationship was found also between PCIN and hypodiploidy ( p = 0.0001), but not with deletion of chromosome 13. This study emphasises the relationship between PCIN, an easy-to-ascertain marker of intrinsic malignancy of the tumour process, and adverse prognosis.

The Effect of Clay Content on PMMA-Clay Nanocomposite Foams

In this study the CO2 sorption at 45 °C in PMMA nanocomposite films containing 2 wt.% of nanoclay has been measured using an in-situ gravimetric technique. The films examined were prepared by compression moulding material obtained by dry-blend and solvent co-precipitation techniques. The CO2 diffusion coefficients were found to be higher for the dry-blended nanocomposite due to the larger agglomerations of the organoclay agglomerations, which prevented the polymer chains from fully wetting and intercalating the clay particles. The Tg-p profile for PMMA nanocomposite containing 2 wt.% nanoclay in the presence of CO2 was also measured using high-pressure DSC. The glass transition phase envelope was shifted vertically by approximately 10 °C when compared to the value reported in the literature for neat PMMA. This result suggests that the nanoclay affects the plasticization behaviour of PMMA under high-pressure CO2 conditions. The cellular morphologies obtained for these PMMA nanocomposite foams produced by batch processing with subcritical CO2 are strongly dependent upon the clay content and the dispersion of the nanoclay in the material. In the case of intercalated nanocomposites, most clay particles exist as agglomerated stacks of silicate sheets. On foaming the cells tend to form around the clay particles causing either irregular-shaped cells or layers to be produced. As a result, the cell density increases and the mean cell size decreases in the foamed nanocomposite on increasing the nanoclay content. Accordingly, the resulting cell structures are highly non-uniform and show large variations in cellular morphologies throughout the foam.

Effect of auxins on Karwinskia humboldtiana root cultures

Effects of auxins (IAA, IBA and NAA) on K. humboldtiana root culture cultivated in 16-h photoperiod or in dark have been observed. Light affected positively the production of biomass when cultivated on medium supplemented with NAA in 10 and 25 μ mol −1 concentrations. In the presence of IAA and IBA these values were significantly lower. The growth dynamics of root cultures depended on the auxin used. The best adventitious roots elongation and lateral roots induction on media supplemented with IBA has been ascertained. Morphological and anatomical differences in dependence on auxin used were observed. NAA supported the formation of huge callus-like mass besides mostly very short roots, especially under the light. Similarly IAA induced short roots, and IBA seems to be the most effective substance for the root elongation in this model system. NAA induced roots with larger diameter under the light compared with the other two auxins used. The reason is in the different anatomical structure of roots which was characterized by higher number of cell layers and large intercellulars in the cortex. The shape of cortical cells in the presence of IBA depended on the light conditions. Isodiametric cortical cells were present in roots cultivated in 16-h photoperiod, irregularly-shaped cells in the dark. The effect of light conditions was the smallest in the case of roots grown on IAA enriched media.

Effects of cell shape and cell wall thickness variations on the elastic properties of two-dimensional cellular solids

The Voronoi tessellation technique and the finite element method are utilized to investigate the microstructure-property relations of two-dimensional cellular solids having irregular cell shapes and non-uniform cell wall thickness. Twenty finite element models are constructed for each type of honeycomb samples (specimens) to obtain the mean values and standard deviations of the effective elastic properties. Spatially periodic boundary conditions are applied to each specimen containing 360 complete cells. The simulation results indicate that the elastic moduli increase as cell shapes become more irregular, but decrease as cell wall thickness gets less uniform. The Poisson’s ratios are insignificantly affected by the presence of these two types of imperfections. The effect of the interaction between the co-existing cell shape and cell wall thickness imperfections on the elastic moduli is found to be weak. Bending remains as the dominant deformation mechanism in a loaded honeycomb having irregular cell shapes and/or non-uniform cell wall thickness. In addition, it is revealed that such imperfect honeycombs can be regarded as isotropic.

Intra-membrane ligand diffusion and cell shape modulate juxtacrine patterning

A key problem in developmental biology is how pattern and planar polarity are transmitted in epithelial structures. Examples include Drosophila neuronal differentiation, ommatidia formation in the compound eye, and wing hair polarization. A key component for the generation of such patterns is direct cell–cell signalling by transmembrane ligands, called juxtacrine signalling. Previous models for this mode of communication have considered homogeneous distributions in the cell membrane, and the role of polarity has been largely ignored. In this paper we determine the role of inhomogeneous protein and receptor distributions in juxtacrine signalling. We explicitly include individual membrane segments, diffusive transport of proteins and receptors between these segments, and production terms with a combination of local and global responses to ligand binding. Our analysis shows that intra-membrane ligand transport is vital for the generation of long wavelength patterns. Moreover, with no ligand transport, there is no pattern formation for lateral induction, a process in which receptor activation up-regulates ligand production. Biased production of ligand also modulates patterning bifurcations and predicted wavelengths. In addition, biased ligand and receptor trafficking can lead to regular polarity across a lattice, in which each cell has the same orientation—directly analogous to patterns of hairs in the Drosophila wing. We confirm the trends in pattern wavelengths previously observed for patterns with cellular homogeneity—lateral inhibition tends to give short-range patterns, while lateral induction can give patterns with much longer wavelengths. Moreover, the original model can be recovered if intra-membrane bound receptor diffusion is included and rapid equilibriation between the sides is considered. Finally, we consider the role of irregular cell shapes and waves in such networks, including wave propagation past clones of non-signalling cells.

Induction of GADD gene expression by phenethylisothiocyanate in human colon adenocarcinoma cells

Phenethylisothiocyanate (PEITC), a potential cancer chemopreventive agent, induces colon cancer cell death, but the mechanism is not entirely clear. Therefore, the aim of this study was to further clarify the molecular effects of PEITC in causing death of human colon adenocarcinoma cells. When incubated with PEITC, HCT-116 colonocytes showed morphological features characteristic of apoptosis, such as irregular cell shape, translocation of plasma membrane phosphatidylserine, and also chromatin condensation and fragmentation. These changes occurred after single-strand breaks in DNA were detected, suggesting that PEITC induced irreparable DNA damage, which in turn triggered the process of apoptosis. DNA macroarray analysis of a selected small cluster of apoptosis-related genes revealed noticeably higher expression of only GADD45, which was confirmed by gene-specific relative RT-PCR analysis. This led to investigation of other GADD gene members possibly affected by PEITC. Whereas GADD34 mRNA expression increased just slightly, there was an appreciable elevation of the mRNA for GADD153, which is recognized as a pro-apoptotic gene. The effect of PEITC on GADD153 was attenuated by either actinomycin D or N-acetylcysteine, suggesting that PEITC-induced upregulation of GADD153 mRNA expression was partly at the level of transcriptional activation involving reactive oxygen species. Additionally, PEITC-induced upregulation of GADD153 mRNA expression did not appear to require p53, based on the observation that PEITC also increased GADD153 mRNA expression in HCT-15 colonocytes, which are known to express mutant p53. These findings suggest that PEITC creates an oxidative cellular environment that induces DNA damage and GADD153 gene activation, which in turn helps trigger apoptosis.

"Primed" human neutrophils on a standard peripheral blood smear.

In the two-event model of end organ injury the first event is the priming of circulating neutrophils (PMNs); the second stressor activates these primed PMNs with subsequent indiscriminate release of cytotoxic reactive oxygen metabolites and proteases into the PMNendothelial microenvironment. After both trauma and cardiopulmonary bypass, circulating PMNs are primed for superoxide release. The first “priming” event in the development of multiple organ failure can be detected with standard light microscopic (100 ) evaluation of routine peripheral blood smears (Wright-Giemsa stain). The resting neutrophil (A) exhibits a smooth, round cell shape with uniform cytoplasmic granularity. After priming (B), with 2 M platelet activating factor, PMNs lose their resting morphology and exhibit an irregular cell shape and membrane, toxic granulation with polarized granularity (white arrowhead), and cytoplasmic vacuolization (black arrowhead).

Unusual cytological patterns of microsporogenesis in Brachiaria decumbens: abnormalities in spindle and defective cytokinesis causing precocious cellularization.

Cytogenetic studies carried out in the tetraploid accession BRA001068 of Brachiaria decumbens, also known as cv. Basilisk, revealed an unusual pattern of microsporogenesis. The spindle in metaphase I and anaphase I became heavily stained with propionic carmine. In telophase I, the interzonal microtubules continued to be intensely stained, and during the phragmoplast formation the fibers were pushed to the cell wall, persisting until prophase II, even after cytokinesis. Due to its tetraploid condition, the accession presented many cells with precocious chromosome migration to the poles in metaphase I and laggards in anaphase I that gave rise to micronuclei in telophase I. While in other polyploid accessions of Brachiaria micronuclei remained in this condition until the second cytokinesis, the micronuclei in this accession organized their own spindle in the second division. In several microsporocytes, the micronuclei with their minispindle were divided further into microcytes by additional cytokinesis. Some curious planes of cytokinesis were found in some cells, with partitioning of cytoplasm into cells of irregular shape. The result consisted of a high frequency of abnormal products of meiosis. Quadrivalents were observed in diakinesis at low frequency, which suggests a segmental allotetraploid and the inability of both genomes to co-ordinate their activities, leading to multiple spindle and precocious cellularization. In spite of abnormal meiotic products reducing pollen fertility, seed production was normal. Enough normal pollen was available to fertilize the central-cell nucleus of the embryo sac and produce normal endosperm in this pseudogamous aposporous apomictic accession.

Repeated fasting stress causes activation of mitogen-activated protein kinases (ERK/JNK) in rat liver

Mitogen-activated protein kinases (MAPK)–signaling pathways play key roles in cytoplasmic-nuclear signal transmission in response to various extracellular stimuli. In this study, we investigated the effect of repeated fasting stress on activation of the 3 members of the MAPK family, the extracellular signal-regulated kinase (ERK), the c-Jun NH 2-terminal kinase (JNK), and the p38 mitogen-activated protein kinase (p38 kinase), in rat liver. Immunecomplex kinase assays showed that ERK and JNK were significantly activated in the liver extract from fasted rats whereas p38 kinase showed no activation. In an immunohistochemical study, the phosphorylated and activated form of ERK (p-ERK) was abundantly expressed in pericentral hepatocytes of fasted liver compared with those of the control. On the other hand, the phosphorylated and activated form of JNK (p-JNK) was highly expressed in irregular-shaped cells along the sinusoidal lining of fasted liver. A double immunofluorescent study to identify p-JNK immunoreactive cells revealed them to be Kupffer cells, which are the resident hepatic macrophages. In conclusion, ERK and JNK are selectively activated in distinct cell types of rat liver by repeated fasting stress. (H EPATOLOGY 2002;36:72-80.)