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Nucleation-catalysis-kinetics analysis under dynamic conditions.

Even at large undercooling, nucleation during solidification is usually controlled by a heterogeneous process. A number of models have been devised to describe the catalytic potency of various potential nucleation sites. Most common analysis models are based upon continuum descriptions that neglect the atomistic structural features that are important at the critical nucleus size scale. Even when surface structures such as steps are considered, they are treated as static stationary features in catalysis models. At the same time, nucleation is recognized as a dynamic process. Experimental strategies to explore some aspects of nucleation dynamics have been developed based upon the droplet method. In droplet dispersions, heterogeneous catalysis can be examined in two-phase liquid-solid mixtures with well-characterized morphologies to reveal surface structure effects and catalyst dynamics.

Biophysical Phenomena↗

Wigglesworthia gen. nov. and Wigglesworthia glossinidia sp. nov., taxa consisting of the mycetocyte-associated, primary endosymbionts of tsetse flies.

The primary endosymbionts (P-endosymbionts) of tsetse flies (Diptera: Glossinidae) are harbored inside specialized cells (mycetocytes) in the anterior region of the gut, and these specialized cells form a white, U-shaped organelle called mycetome. The P-endosymbionts of five tsetse fly species belonging to the Glossinidae have been characterized morphologically, and their 16S ribosomal DNA sequences have been determined for phylogenetic analysis. These organisms were found to belong to a distinct lineage related to the family Enterobacteriaceae in the gamma subdivision of Proteobacteria, which includes the secondary endosymbionts of various insects and Escherichia coli. These bacteria are also related to the P-endosymbionts of aphids, Buchnera aphidicola. Signature sequences in the 16S ribosomal DNA and genomic organizational differences which distinguish the tsetse fly P-endosymbionts from members of the Enterobacteriaceae and from the genus Buchnera are described in this paper. I propose that the P-endosymbionts of tsetse flies should be classified in a new genus, the genus Wigglesworthia, and a new species, Wigglesworthia glossinidia. The P-endosymbiont found in the mycetocytes of Glossina morsitans morsitans is designated the type strain of this species.

Animals↗

New type B retrovirus isolates associated with kinetochores and centrioles of the host cell.

Two new virus isolates, M432 and M832, obtained from the Southeast Asian mouse have been characterized morphologically with respect to their composition and intracellular assembly. The mature virions resemble in certain respects the type B murine retroviruses. The new isolates, however, have an intracellular precursor, a type A particle, closely associated with the mitotic apparatus. The intracellular transport of the type A particles to the cell surface, where they are released by budding, is closely associated with the microtubule system of the cell.

Cell Line↗

Down-regulation of integrin alpha 1/beta 1 expression and association with cell rounding in human cytomegalovirus-infected fibroblasts.

Human cytomegalovirus (HCMV) causes a c.p.e. characterized by rounding of the infected cell. Since interactions with the extracellular matrix may be involved in the cell rounding, we have analysed the expression of integrins, which are the main cell surface receptors involved in cell-substrate adhesion and spreading. By FACS analysis, a selective decrease in cell surface expression of alpha 1/beta 1 integrin was observed in HCMV-infected fibroblasts. This decrease coincides with cell rounding. Immunoprecipitation studies and FACS analysis of permeabilized cells have further demonstrated that total levels of this integrin are decreased in infected cells, suggesting that the reduction in cell surface alpha 1/beta 1 integrin is not due to a defect in transport to the surface. Furthermore, we have ruled out the possibility that the observed decrease in alpha 1/beta 1 expression is caused by a cytokine released from the infected cells by showing that the reduction is abolished by inactivating the HCMV with u.v. irradiation, and that conditioned medium from HCMV-infected cells has no effect on expression of alpha 1/beta 1 integrin in uninfected cells. Concomitant with the reduction in alpha 1/beta 1 levels, the HCMV-infected fibroblasts show a reduced ability to adhere to laminin and collagen IV. Taken together the data indicate that de novo synthesis of HCMV protein(s) causes a decreased assembly/expression of alpha 1/beta 1 integrin, coincident with the well characterized morphological alterations of the infected cell.

Cell Line↗

Nocardia takedensis sp. nov., isolated from moat sediment and scumming activated sludge.

Chemotaxonomic and morphological characterization of two actinomycete strains, MS1-3T and AS4-2, respectively isolated from moat sediment and scumming activated sludge, was carried out. This characterization clearly demonstrated that strains MS1-3T and AS4-2 belong to the genus Nocardia. 16S rRNA gene sequencing studies showed that these isolates are most closely related to Nocardia beijingensis (98.1-98.3 % similarity), Nocardia brasiliensis (97.9-98.0 %) and Nocardia tenerifensis (97.8-97.9 %). However, the results of DNA-DNA hybridizations and physiological and biochemical tests showed that strains MS1-3T and AS4-2 could be differentiated from their closest phylogenetic relatives both genotypically and phenotypically. It is proposed that the two isolates be classified as representatives of a novel species of Nocardia, Nocardia takedensis sp. nov. The type strain is MS1-3T (=NBRC 100417T=DSM 44801T); AS4-2 (=NBRC 100418=DSM 44802) is a reference strain.

Bacterial Typing Techniques↗

Azospirillum melinis sp. nov., a group of diazotrophs isolated from tropical molasses grass.

Fifteen bacterial strains isolated from molasses grass (Melinis minutiflora Beauv.) were identified as nitrogen-fixers by using the acetylene-reduction assay and PCR amplification of nifH gene fragments. These strains were classified as a unique group by insertion sequence-PCR fingerprinting, SDS-PAGE protein patterns, DNA-DNA hybridization, 16S rRNA gene sequencing and morphological characterization. Phylogenetic analysis of the 16S rRNA gene indicated that these diazotrophic strains belonged to the genus Azospirillum and were closely related to Azospirillum lipoferum (with 97.5 % similarity). In all the analyses, including in addition phenotypic characterization using Biolog MicroPlates and comparison of cellular fatty acids, this novel group was found to be different from the most closely related species, Azospirillum lipoferum. Based on these data, a novel species, Azospirillum melinis sp. nov., is proposed for these endophytic diazotrophs of M. minutiflora, with TMCY 0552(T) (=CCBAU 5106001(T) = LMG 23364(T) = CGMCC 1.5340(T)) as the type strain.

Acetylene↗

Kineococcus marinus sp. nov., isolated from marine sediment of the coast of Jeju, Korea.

A novel marine actinomycete, designated strain KST3-3(T), which was isolated from a sediment sample of the coast of Jeju, Korea, was subjected to a polyphasic taxonomic characterization. The organism was characterized morphologically by the formation of motile, coccoid cells. A phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that the organism was related to the genera Kineosporia and Kineococcus, and members of the suborder Frankineae, and occupied the deepest branch outside a taxon encompassing members of the genus Kineococcus. The organism showed relatively low levels of 16S rRNA gene sequence similarity to members of the genera Kineococcus (93.0-93.4 %) and Kineosporia (93.1-93.8 %). The morphological and chemotaxonomic characteristics, albeit with a slightly higher level of sequence similarity to members of the genus Kineosporia, supported its classification within the genus Kineococcus. On the basis of the polyphasic data presented, it was evident that the organism should be assigned to a novel species of the genus Kineococcus, for which the name Kineococcus marinus sp. nov. is proposed. The type strain is strain KST3-3(T) (=KCCM 42250(T) = NRRL B-24439(T)).

Actinomycetales↗

Diversity of stx2 converting bacteriophages induced from Shiga-toxin-producing Escherichia coli strains isolated from cattle.

The presence of bacteriophages encoding Shiga toxin 2 (stx(2) phages) was analysed in 168 strains of Shiga-toxin-producing Escherichia coli (STEC) isolated from cattle. Following mitomycin C induction, strains carrying stx(2) phages were screened by plaque blot and hybridization with an stx(2)A-probe. In the stx(2)-phage-carrying strains, the amounts of phage production, phage DNA extracted and Stx(2) produced after induction were assessed. The induced stx(2) phages were characterized morphologically and genetically. Assays to obtain lysogens from different strains were also carried out and phages induced from the lysogens were compared with those induced from the STEC isolates. Results indicated that 18 % of the strains carried an inducible stx(2) phage. Most of them showed a direct relationship between phage induction and toxin production. Each strain carried only one inducible stx(2) phage, although a few strains had two copies of the stx(2) in the chromosome. The stx(2) phages showed diverse morphology and a wide variability in their genome. Assays to obtain lysogens showed that not all the phages were transduced with the same frequency and only six lysogens were obtained. Phages in the lysogens were the same as those induced from their respective initial STEC host strains, although the induction and relative toxin production of the lysogens varied. Most phages carried the stx(2) gene, while a few carried stx(2) variants. Infectivity of the phages depended on the different hosts, although O157 : H7 was preferentially infected by phages induced from O157 strains. The results show that inducible stx(2) phages are common among STEC of animal origin and that they may enhance the spread of stx(2).

Amino Acid Sequence↗

Flow correlated percolation during vascular remodeling in growing tumors.

A theoretical model based on the molecular interactions between a growing tumor and a dynamically evolving blood vessel network describes the transformation of the regular vasculature in normal tissues into a highly inhomogeneous tumor specific capillary network. The emerging morphology, characterized by the compartmentalization of the tumor into several regions differing in vessel density, diameter, and necrosis, is in accordance with experimental data for human melanoma. Vessel collapse due to a combination of severely reduced blood flow and solid stress exerted by the tumor leads to a correlated percolation process that is driven towards criticality by the mechanism of hydrodynamic vessel stabilization.

Blood Vessels↗

An Arabidopsis mutant defective in jasmonate response is allelic to the auxin-signaling mutant axr1.

A screen for Arabidopsis mutants that were insensitive to methyl jasmonate (MeJA) in an assay for seedling root growth yielded only alleles of previously isolated mutants jar1 and coi1, with one exception. Mapping of the locus and morphological characterization of the new mutant suggested it might be allelic to axr1, which had not previously been reported to show resistance to MeJA. The F(1) from a cross of the new mutant with axr1-3 did not show complementation, confirming that these are the same genes. The new allele is called axr1-24. In addition to MeJA and indole-3-acetic acid (IAA), axr1-24 had decreased sensitivity to 1-aminocyclopropane-1-carboxylic acid, 6-benzylamino-purine, epi-brassinolide, and abscisic acid. Both axr1-24 and the previously characterized axr1-3 allele were shown to be susceptible to the opportunistic pathogen Pythium irregulare, a trait found in other jasmonate response mutants, including jar1-1. The double mutant jar1-1/axr1-3 was more resistant to inhibition of root growth by MeJA and was more susceptible to P. irregulare infection than either single mutant, suggesting these genes might act in independent response pathways. In contrast, resistance to IAA in the double mutant was not different from axr1-3. Northern-blot analysis showed that IAA induced the jasmonate-responsive lipoxygenase 2, AOS, and AtVSP gene transcripts and induction was strongly impaired in axr1-3. However, transcript induction by MeJA was only minimally affected in axr1-3. This study demonstrates that in addition to auxin signaling, the AXR1 locus is involved in MeJA response, providing a mechanistic link between jasmonate and auxin-signaling pathways.

Abscisic Acid↗

Heterologous expression of Arabidopsis phytochrome B in transgenic potato influences photosynthetic performance and tuber development.

Transgenic potato (Solanum tuberosum) plants expressing Arabidopsis phytochrome B were characterized morphologically and physiologically under white light in a greenhouse to explore their potential for improved photosynthesis and higher tuber yields. As expected, overexpression of functional phytochrome B caused pleiotropic effects such as semidwarfism, decreased apical dominance, a higher number of smaller but thicker leaves, and increased pigmentation. Because of increased numbers of chloroplasts in elongated palisade cells, photosynthesis per leaf area and in each individual plant increased. In addition, photosynthesis was less sensitive to photoinactivation under prolonged light stress. The beginning of senescence was not delayed, but deceleration of chlorophyll degradation extended the lifetime of photosynthetically active plants. Both the higher photosynthetic performance and the longer lifespan of the transgenic plants allowed greater biomass production, resulting in extended underground organs with increased tuber yields.

Arabidopsis↗

Complex permittivity measurement as a new noninvasive tool for monitoring in vitro tissue engineering and cell signature through the detection of cell proliferation, differentiation, and pretissue formation.

In in vitro tissue engineering, microporous scaffolds are commonly used to promote cell proliferation and differentiation in three-dimensional structures. Classic measurement methods are particularly time consuming, difficult to handle, and destructive. In this study, a new nondestructive method based on complex permittivity measurement (CPM) is proposed to monitor and track the osteoblast and macrophage differentiation through their morphological variation upon cell attachment and proliferation inside the microporous scaffolds. CPM is performed using a vector network analyzer and a dielectric probe under sterile conditions in a laminar-flow hood. A suitable effective medium approximation (EMA) is applied to fit the data in order to extract the parameters of the different constituents. Our data show that the EMA depolarization factor can be monitored to assess the variation of cell morphology characterizing cell attachment. Discrimination between two batches of scaffolds seeded, respectively, with 2 million and 1 million osteoblast cells is possible; the ratio of their CPM-derived cell volume fractions is in agreement with the ratio of their cell seeding numbers. In addition, cell proliferation inside scaffolds seeded with osteoblasts cultured in alpha minimum essential medium and inside scaffolds seeded with osteoblasts cultured in alpha minimum essential medium supplemented to induce the formation of extracellular matrix is monitored via CPM over several days. CPM-determined cell volume fraction is compared to DNA assay cell counts. Extracellular matrix formation and cell presence was confirmed by scanning electron microscopy. A set of three signature parameters (epsilon'mem, epsilon'cyt, kappa'cyt) characteristic of cell line is extracted from CPM. Distinct signatures are recorded for osteoblasts and macrophages, thus confirming the ability of CPM to discriminate between different cell types. This study demonstrates the potential of CPM as a diagnostic tool to monitor quickly and noninvasively cell growth and differentiation inside microporous scaffolds. Our findings suggest that the use of CPM could be extended to many biomedical applications, such as drug detection and automation of tissue and bacterial cultures in bioreactors.

3T3 Cells↗

Oxidative phosphorylation in rat oral mucosal mitochondria.

Oral mucosal mitochondria were isolated and characterized morphologically by electron microscopy. Polarographic measurements were made of respiration and oxidative phosphorylation in the mitochondrial preparations. ADP:O ratios approaching or slightly exceeding the theoretical maxima and stabilized respiratory control ratios were achieved with malate + glutamate, succinate and ascorbate-N,N,N1N1 tetramethyl-p-phenylenediamine (TMPD) as substrates. Inhibition by rotenone, antimycin A, azide, and cyanide established the classical electron transport chain as the major pathway of mitochondrial respiration. Respiration of the oral mucosal mitochondria was stimulated by DNP in the presence of succinate. DNP-stimulated respiration exceeded that observed in the presence of ADP plus Pi and increasing the concentration of DNP progressively inhibited respiration.

Acid Phosphatase↗

The evolution of larval morphology and swimming performance in ascidians.

The complexity of organismal function challenges our ability to understand the evolution of animal locomotion. To meet this challenge, we used a combination of biomechanics, phylogenetic comparative analyses, and theoretical morphology to examine evolutionary changes in body shape and how those changes affected swimming performance in ascidian larvae. Results of phylogenetic comparative analyses suggest that coloniality evolved at least three times among ascidians and that colonial species have a convergent larval morphology characterized by a large trunk volume and shorter tail length in proportion to the trunk. To explore the functional significance of this evolutionary change, we first verified the accuracy of a mathematical model of swimming biomechanics in a solitary (C. intestinalis) and a colonial (D. occidentalis) species and then ran numerous simulations of the model that varied in tail length and trunk volume. The results of these simulations were used to construct landscapes of speed and cost of transport predictions within a trunk volume/tail length morphospace. Our results suggest that the reduction of proportionate tail length in colonial species resulted in improved energetic economy of swimming. The increase in the size of larvae with the origin of coloniality facilitated faster swimming with negligible energetic cost, but may have required a reduction in adult fecundity. Therefore, the evolution of ascidians appears to be influenced by a trade-off between the fecundity of the adult stage and the swimming performance of larvae.

Animals↗

Sandflies of the Phlebotomus perniciosus complex: mitochondrial introgression and a new sibling species of P. longicuspis in the Moroccan Rif.

The bloodsucking adult females of Phlebotomus perniciosus Newstead and P. longicuspis Nitzulescu (Diptera: Psychodidae) are important vectors of the protozoan Leishmania infantum Nicolle (Kinetoplastida: Trypanosomatidae) in western Mediterranean countries. The species status of the two phlebotomine sandflies was assessed, along with the epidemiological implications. Individual sandflies from three Moroccan Rif populations were characterized morphologically, isoenzymatically (by the isoelectrofocusing of alleles at the polymorphic enzyme loci of HK, GPI and PGM), and by comparative DNA sequence analysis of a fragment of mitochondrial Cytochrome b (mtDNA). By reference to the character profiles of specimens from other locations, including southern Spain and the type-locality countries, the Moroccan flies were placed in three lineages: first, the lineage of P. perniciosus, which contained two mtDNA sublineages, one (pnt) widely distributed and associated with the morphology of the male types from Malta, and the other (pna) associated with a P. longicuspis-like male morphology; second, the lineage of P. longicuspis sensu stricto, including typical forms from Tunisia; and third, a new sibling species of P. longicuspis. The mtDNA sublineage (pnt) of typical P. perniciosus was also found in some P. longicuspis from Morocco, indicating interspecific hybridization. The typical race of P. perniciosus occurs in Italy as well as in Malta, Tunisia and Morocco. It is replaced in southern Spain by the Iberian race (with the pni mtDNA sublineage). The discovery of interspecific gene introgression and a new sibling species mean that previous records of the two morphospecies do not necessarily reflect their true vectorial roles or geographical and ecological distributions.

Alleles↗

The protean spectrum of non-Hodgkin lymphomas with prominent involvement of subcutaneous fat.

BACKGROUND: Subcutaneous T-cell lymphoma (STCL) represents a controversial entity and a confused concept in the field of cutaneous T-cell lymphomas (CTCLs). Recently, alpha/beta+/CD8+ STCL has been recognized by the new World Health Organization (WHO)-European Organization for Research and Treatment of Cancer (EORTC) classification of primary cutaneous lymphomas as a distinct entity in the group of CTCLs. OBSERVATIONS: We reviewed a series of 53 biopsies from 26 patients (F : M = 19:7; median age: 48; range 18-87) of cutaneous B- and T-cell lymphomas characterized by prominent involvement of the subcutaneous tissue. We could classify our cases according to the following seven categories--(i) STCL: n = 16; (ii) extranodal NK/T-cell lymphoma, nasal type: n = 2; (iii) cutaneous gamma/delta T-cell lymphoma: n = 2; (iv) anaplastic CD30+ large T-cell lymphoma: n = 1; (v) diffuse large B-cell lymphoma, secondary cutaneous: n = 3; (vi) lymphoplasmacytic lymphoma, secondary cutaneous: n = 1; (vii) specific cutaneous manifestations of myelogenous leukemia: n = 1. CONCLUSIONS: We demonstrated the protean nature of lymphomas with prominent involvement of the subcutaneous fat tissues. The term STCL should be restricted to a homogeneous group of cases characterized morphologically by an exclusive involvement of subcutaneous tissues, immunohistochemically by a T-cytotoxic alpha/beta phenotype, and biologically by a relatively good prognosis.

Adipose Tissue↗

Morphological properties of a human intestinal spirochete first isolated from a patient with diarrhea in Japan.

A human intestinal spirochete isolated from a rectal biopsy specimen was morphologically characterized. The isolate was comma-shaped, 3-6 microm in length, 0.2 micro m in diameter and had tapered ends. The surface layer, external to the outer envelope, was amorphous. Four string-like periplasmic flagella with a diameter of 20 nm were presented at each end of the SDS-treated cells. Thin sections of the bacterial cell revealed a high-density cytoplasmic membrane and flagella in the periplasmic space between the cytoplasmic membrane and the outer envelope. Three segments of equal length were observed in some of the cells, while other cells were bi-segmented and more frequently observed.

Biopsy↗

Microvascular changes in the stroma of human colorectal carcinomas: ultrastructural histochemical study.

Ultrastructural histochemical studies were performed using Ulex europaeus agglutinin-I lectin (UEA-I) and anti-endothelial monoclonal antibody BMA 120 in order to morphologically characterize the microvasculature of human colorectal carcinomas. In the normal mucosa, UEA-I and BMA 120 were bound to luminal plasma membrane of endothelial cells, usually continuously. Capillaries in the stroma of invasive adenocarcinomas showed remarkable structural changes such as swelling of endothelial cells with well-developed cell organelles and narrowing of the lumen. Reaction products for UEA-I and BMA 120 were both observed along the luminal plasma membrane of endothelial cells, usually discontinuously, partially retaining the features of normal capillaries. Furthermore, we have confirmed an occurrence of solid capillary buds composed of a strand of large endothelial cells with a trace of lumen. They showed almost no reactivity to UEA-I and BMA 120. Conventional electron microscopy revealed that they were present frequently in invasive carcinomas, but infrequently in intramucosal carcinomas and inflammatory lesions. Our results suggest that the stroma of invasive colorectal carcinomas abounds in immature capillaries and their precursors, which may indicate active tumor-induced angiogenesis.

Antibodies, Monoclonal↗