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Microscopic Observations (microscopic + observation)
Kinds of Microscopic Observations Selected AbstractsVisible and FTIR Microscopic Observation of Bisthiourea Ionophore Aggregates in Ion-Selective Electrode MembranesELECTROANALYSIS, Issue 22 2005Katherine Abstract Since conventional response models for ionophore-based ISEs are based on the assumption of a homogeneous membrane phase, they cannot accurately predict the response of membranes containing self-aggregating ionophores. However, meaningful conclusions about the relationship between ionophore structure and potentiometric responses can only be drawn if ionophore aggregation is properly recognized. This study demonstrates that dark field visible microscopy and FTIR microspectroscopy are valuable tools for the observation of such ionophore self-aggregation and, thereby, the development of new ionophore-based ISEs. Sulfate selective electrodes with solvent polymeric membranes containing bisthiourea ionophores that differ only by peripheral nonpolar substituents were shown to exhibit very different interferences from the sample pH. On one hand, optimized electrodes based on an ionophore with a phenyl substituent on each thiourea group (1) do not respond to pH at all and function well as sulfate-selective electrodes. On the other hand, membranes containing a more lipophilic ionophore with two additional hexyl-substituted adamantyl groups (2) exhibit severe pH interference at pH values as low as pH,5. The observation of membranes containing ionophore 2 with dark field visible microscopy and FTIR microspectroscopy shows supramolecular aggregation, and explains the startling difference between the potentiometric responses of the two types of electrodes. [source] Atomic Force Microscopic Observations on the Crystalline Morphology of Poly(ethylene naphthalate)/Clay NanocompositesMACROMOLECULAR CHEMISTRY AND PHYSICS, Issue 16 2007Yang Choo Chua Abstract Atomic force microscopic observations on an isothermally crystallized poly(ethylene naphthalate) (PEN)/clay nanocomposite suggest that the presence of nanoclay alters the lamellar organization in PEN mainly in three ways: 1) physically blocking the crystal growth front and creating wide amorphous regions within the spherulites, which may then be filled by secondary lamellae branching out from the primary lamellae of the same spherulite, or primary lamellae developed from other nearby nucleating centers; 2) inducing random twisting of lamellae; and 3) causing irregular crystallite growth fronts, with the protrusion of some leading lamellae. In particular, the physical hindrance imposed by clay tends to be more prevalent for lamellae that grow roughly perpendicular to the clay long axes. This may give rise to an anisotropic crystalline morphology if the clay layers exhibit a preferred orientation induced by flow. [source] EX VIVO CASE STUDY OF ENDOCYTOSCOPY IN SUPERFICIAL ESOPHAGEAL CANCERDIGESTIVE ENDOSCOPY, Issue 2007Mototsugu Kato Microscopic observation at the cellular level using endocytoscopy was obtained in surface mucosa of the gastrointestinal tract. This paper describes ex vivo images for endoscopically resected specimens of superficial esophageal cancer using endocytoscopy with methylene blue staining. The endocytoscopy images of cancerous and non-cancerous sites corresponded generally with horizontal histological images. The pattern of the cellular arrangement and the size and shape of cells were similar between endocytoscopy and horizontal histological imaging. Endocytoscopy is an effective tool for diagnosis of esophageal cancer. [source] Halophilic archaea in the human intestinal mucosaENVIRONMENTAL MICROBIOLOGY, Issue 9 2010Andrew P. A. Oxley Summary The human gastrointestinal tract microbiota, despite its key roles in health and disease, remains a diverse, variable and poorly understood entity. Current surveys reveal a multitude of undefined bacterial taxa and a low diversity of methanogenic archaea. In an analysis of the microbiota in colonic mucosal biopsies from patients with inflammatory bowel disease we found 16S rDNA sequences representing a phylogenetically rich diversity of halophilic archaea from the Halobacteriaceae (haloarchaea), including novel phylotypes. As the human colon is not considered a salty environment and haloarchaea are described as extreme halophiles, we evaluated and further discarded the possibility that these sequences originated from pre-colonoscopy saline lavage solutions. Furthermore, aerobic enrichment cultures prepared from a patient biopsy at low salinity (2.5% NaCl) yielded haloarchaeal sequence types. Microscopic observation after fluorescence in situ hybridization provided evidence of the presence of viable archaeal cells in these cultures. These results prove the survival of haloarchaea in the digestive system and suggest that they may be members of the mucosal microbiota, even if present in low numbers in comparison with methanogenic archaea. Investigation of a potential physiological basis of this association may lead to new insights into gastrointestinal health and disease. [source] Anthracene photoinduced toxicity to plhc-1 cell line (Poeciliopsis lucida) and the role of lipid peroxidation in toxicityENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 11 2000Jonghoon Choi Abstract Many polycyclic aromatic hydrocarbons (PAHs) are acutely toxic to fish and other aquatic organisms in the presence of solar ultraviolet radiation (SUVR) of environmentally realistic intensities. In the present study, the photoinduced toxicity of a PAH (anthracene; ANT) to topminnow hepatoma cell line (PLHC-1) was assessed. After the toxicity was characterized, the role of lipid peroxidation in PAH photoinduced toxicity was examined by measuring lipid peroxidation products and by assessing the effect of lipid peroxidation antagonist (Trolox) treatment. In cytotoxicity tests using two assays (MTT, neutral red), the SUVR/ANT treatment elicited toxicity to PLHC-1 cells in a concentration- and SUVR (exposure duration and intensity)-dependent pattern. As found in previous organism-level studies, no significant cytotoxicity was observed in the cells exposed either to fluorescent light/ANT or to SUVR only. The SUVR/ANT treatment elicited the lipid peroxidation process and Trolox pretreatment significantly reduced SUVR/ANT-induced cell mortality. Microscopic observation showed that Trolox pretreatment relieved the SUVR/ANT-inflicted damage, such as cell shrinkage and membrane disruption. Together with a recent finding in our lab that increased production of superoxide anion and a lipid peroxidation product (malondialdehyde) was found in SUVR/ANT-treated fish microsomes, the present study suggests that reactive oxygen radical-induced lipid peroxidation is an important factor in PAH photoinduced toxicity to fish. [source] Microscopic observation of aerobic granulation in sequential aerobic sludge blanket reactorJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2001J.-H. Tay Aims: This paper attempts to provide visual evidence of how aerobic granulation evolves in sequential aerobic sludge blanket reactors. Methods and Results: A series of experiments were conducted in two column-type sequential aerobic sludge reactors fed with glucose and acetate as sole carbon source, respectively. The evolution of aerobic granulation was monitored using image analysis and optical and scanning electron microscopy. The results indicated that the formation of aerobic granules was a gradual process from seed sludge to compact aggregates, further to granular sludge and finally to mature granules with the sequential operation proceeding. Glucose- and acetate-fed granules have comparable characteristics in terms of settling velocity, size, shape, biomass density and microbial activity. However, the microbial diversity of the granules was associated with the carbon source supplied. In this work, an important aerobic starvation phase was identified during sequential operation cycles. It was found that periodical aerobic starvation was an effective trigger for microbial aggregation in the reactor and further strengthened cell,cell interaction to form dense aggregates, which was an essential step of granulation. The periodical starvation-induced aggregates would finally be shaped to granules by hydrodynamic shear and flow. Conclusions: Aerobic granules can be formed within 3 weeks in the systems. The periodical starvation and hydrodynamic conditions would play a crucial role in the granulation process. Significance and Impact of the Study: Aerobic granules have excellent physical characteristics as compared with conventional activated sludge flocs. This research could be helpful for the development of an aerobic granule-based novel type of reactor for handling high strength organic wastewater. [source] A CALCIUM-DEPENDENT PROTEIN KINASE FUNCTIONS IN WOUND HEALING IN VENTRICARIA VENTRICOSA (CHLOROPHYTA)JOURNAL OF PHYCOLOGY, Issue 6 2000Koh-ichi Sugiyama The cytoplasm around a wound made in the multinucleate unicellular green alga Ventricaria ventricosa ( J. Agardh) Olsen et West formed an aggregation-ring surrounding the wound immediately after injury. A contraction of the ring then brought about wound healing in culture medium containing Ca2+. Involvement of a calcium-dependent protein kinase (CDPK) as a regulator of wound healing was examined using an anti- Dunaliella tertiolecta CDPK antibody. A 52-kDa protein cross-reacting with the antibody was detected by Western blotting. Protein kinases of 60 kDa and 52 kDa, which were markedly activated by Ca2+, and a 40-kDa Ca2+ -independent protein kinase were detected by an in-gel protein kinase assay using myelin basic protein as the substrate. A 52-kDa band with Ca2+ -dependent protein kinase activity was immunoprecipitated from the cytoplasmic extract, indicating that these 52-kDa proteins are identical and possess CDPK activity. Microscopic observation showed that the contraction of the aggregation ring was suppressed by application of the anti-CDPK to the culture medium. A protein kinase inhibitor, K-252a, and the calmodulin inhibitors, calmidazolium and compound 48 / 80, which inhibit CDPK activity, also suppressed the contraction of the aggregation-ring. Immunofluorescence microscopy showed a similar distribution of 52-kDa CDPK to the distribution of f-actin, which was randomly distributed in an intact cell and formed a bundle during wound healing. Further, f-actin was not recruited after injury in the presence of the antibody to CDPK. These results suggest that the 52-kDa CDPK functions as a Ca2+ receptor in wound healing and simultaneously participates in the organization and contraction of f-actin to heal the wound. [source] Live imaging of fluorescent proteins in chordate embryos: From ascidians to miceMICROSCOPY RESEARCH AND TECHNIQUE, Issue 3 2006Yale J. Passamaneck Abstract Although we have advanced in our understanding of the molecular mechanisms intrinsic to the morphogenesis of chordate embryos, the question of how individual developmental events are integrated to generate the final morphological form is still unresolved. Microscopic observation is a pivotal tool in developmental biology, both for determining the normal course of events and for contrasting this with the results of experimental and pathological perturbations. Since embryonic development takes place in three dimensions over time, to fully understand the events required to build an embryo we must investigate embryo morphogenesis in multiple dimensions in situ. Recent advances in the isolation of naturally fluorescent proteins, and the refinement of techniques for in vivo microscopy offer unprecedented opportunities to study the cellular and molecular events within living, intact embryos using optical imaging. These technologies allow direct visual access to complex events as they happen in their native environment, and thus provide greater insights into cell behaviors operating during embryonic development. Since most fluorescent protein probes and modes of data acquisition are common across species, we have chosen the mouse and the ascidian, two model organisms at opposite ends of the chordate clade, to review the use of some of the current genetically-encoded fluorescent proteins and their visualization in vivo in living embryos for the generation of high-resolution imaging data. Microsc. Res. Tech. 69:160,167, 2006. © 2006 Wiley-Liss, Inc. [source] Genetic analysis and gene mapping of a rice recessive male sterile mutantPLANT BREEDING, Issue 3 2010J. B. Chen With 3 figures and 2 tables Abstract Male sterility of rice is one of the major genetic tools used for hybrid rice production. In this study, a spontaneous male sterile mutant, SC-ms-2, was obtained from the F4 progeny of the cross D 297B × Changfeng B. Microscopic observation revealed that the microspores were developed abnormally and the tapetum cells were incrassated during microsporogenesis. Genetic analysis indicated that male sterility of SC-ms-2 was controlled by a single recessive gene. By using bulked segregant analysis on two F2 populations developed from crossing SC-ms-2 with Hua B and ,Nipponbare', this gene was finely mapped between two simple sequence repeat (SSR) markers on chromosome 9, RM24451 and RM7048, with genetic distance of 0.3 cM and 0.6 cM respectively, and the approximate physical distance was 172 kb. Our results showed that this gene was distinguished from all the other male sterility genes in rice reported and it was designated ms92(t), temporally. Moreover, candidate genes in the region of 172kb, including the rice homologue to the Arabidopsis MALE STERILITY1 (MS1) gene, were surveyed and discussed. [source] Microscopic detection of IgY-Fc binding signal in the inner layers of ovarian follicular tissue in quailANIMAL SCIENCE JOURNAL, Issue 5 2010Kohji KITAGUCHI ABSTRACT In avian species, it has been assumed that an Fc receptor in the ovarian follicles mediates immunoglobulin Y (IgY) transport into the yolk. However, no such receptor responsible for IgY has been identified to date. To examine potential IgY binding activity in the entire ovarian follicle, whole-mount sections of quail ovarian follicle were incubated with the Fc fragment of chicken IgY (cIgY). Whole-mount frozen sections of the second largest ovarian follicle were prepared, and then the sections were incubated with digoxigenin-labeled Fc or Fab fragments of cIgY. Microscopic observation revealed that incubation with the cIgY-Fc fragment produced a binding signal in the inner layer of the ovarian follicular tissues, most likely in the granulosa cell layer. However, no such signal was detected when the sections were incubated with cIgY-Fab. Coincubation of the ovarian sections with Alexa488-labeled cIgY-Fc and antiserum raised against ZP1, an envelope protein specifically localized in the perivitelline layer, demonstrated that the source of the Fc binding signals partly coincided with the perivitelline layer. In conclusion, our data suggest that potential IgY binding substances interacting with the Fc domain are present in the inner layers of ovarian follicular tissues, most likely in the granulosa cell layer and/or in the perivitelline layer. [source] Formation of zinc protoporphyrin IX in Parma-like ham without nitrate or nitriteANIMAL SCIENCE JOURNAL, Issue 2 2009Jun-ichi WAKAMATSU ABSTRACT Zinc protoporphyrin IX (ZPP) is a characteristic red pigment in meat products that are manufactured without the addition of a curing agent such as nitrate or nitrite. To examine the effects of impurities such as mineral components in sea salt on the formation of ZPP, we manufactured Parmatype dry-cured hams that were salted with refined salt or sea salt and examined the involvement of oxidation-reduction potential (ORP) in the formation of ZPP. The content of ZPP was increased drastically after 40 weeks. Microscopic observation showed strong fluorescence caused by ZPP muscle fiber after 40 weeks. Conversely, heme content varied considerably during processing. ORP increased during processing. However, there was no obvious difference between ham salted with refined salt and that salted with sea salt. Therefore, it was concluded that impurities in sea salt were not involved in the formation of ZPP. [source] Mode I crack propagation in concrete under fatigue: microscopic observations and modellingINTERNATIONAL JOURNAL FOR NUMERICAL AND ANALYTICAL METHODS IN GEOMECHANICS, Issue 13 2002A. Toumi Abstract In this study, three-point bending tests were carried out on notched beams to investigate mode I crack propagation in plain concrete under fatigue. The first part of the study focused on microscopic observations of the crack growth features. Microscopic observations were made using the replica method associated with scanning electron microscopy (SEM). Observations of fatigue crack growth both on the surface and inside the specimens are presented as a comparison between the observed crack lengths and those estimated by the compliance calibration method. In the second part, a finite element model of mode I crack propagation under fatigue is presented. According to the cohesive crack concept, a cohesive force distribution on the crack at various loading stages is assumed, according to both the stress-crack opening relation worked out by Hordijk (1991; Thesis, Technische Universiteit) and a new proposed relation with hysteresis loop. Finite element computation is used to evaluate the crack extension in the bending beams. Numerical predictions are discussed in comparison with experimental results. Copyright © 2002 John Wiley & Sons, Ltd. [source] Behavior of dense and porous hydroxyapatite implants and tissue response in rat femoral defectsJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2002J.C.T. Andrade Abstract Porous and dense hydroxyapatite cylinders (PHA and DHA) were implanted into cavities produced in rat femora and the sites of implantation were examined at different times over a period of 24 weeks by microradiologic and histological techniques. Microradiographs showed the presence of a layer of trabecular bone around the implants, which became more radiopaque and thinner along the experimental time. The microradiologic methodology used was suitable for the evaluation of the interface between hydroxyapatite and newly formed bone in nondecalcified materials. Microscopic observations showed that young bone grew over the surface of both types of implants after 1 and 2 weeks of surgery and that bone also grew inside PHA implants. Progressive bone absorption was observed in both types of implants after the fourth week. A layer of fibrous tissue was formed in the interface between new bone and DHA. Mature bone with haversian systems surrounded DHA implants and filled the pores of PHA implants throughout the experimental period. The pores of PHA implants were smaller than those commonly reported, which should have been a disadvantage, although it was observed that the extra cellular fluid induced disintegration of the ceramic granules, allowing the gradual growth of bone tissue into the spaces among them, without the interposition of fibrous tissue. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 62: 30,36, 2002 [source] Mineral evolution of a garnet-pyroxenite nodule within eclogite, eastern Sulu ultrahigh-pressure metamorphic terrane, East ChinaJOURNAL OF METAMORPHIC GEOLOGY, Issue 8 2005T. N. YANG Abstract Detailed microtextural observations and bulk chemical analysis were undertaken on a garnet-pyroxenite nodule within retrograde eclogites from the NE Sulu ultrahigh-pressure metamorphic (UHPM) terrane. The results suggest that the protolith was a cumulate from a gabbroic body. The nodule consists primarily of coarse clinopyroxene grains with a very high content of the Ca-Tschermakite molecule. Microscopic observations and back-scattered electron images (BSE) demonstrate a complicated intergrowth of clinopyroxene, garnet and ilmenite, which represents the peak metamorphic assemblage. The primary clinopyroxene grains are armoured with a thin garnet corona up to 0.5 mm wide that forms an interconnected network. Within the clinopyroxene grains, four sets of garnet lamellae are distributed along crystallographic planes; locally, a vermicular intergrowth of garnet and diopside is developed. Besides the garnet, parallel arrays of ilmenite blebs are common within the clinopyroxene. Hydrous minerals such as amphibole, zoisite and titanite formed at later stages, and replaced diopside, garnet and ilmenite respectively. The P,T conditions determined for the formation of the garnet lamellae indicate that the garnet pyroxenite experienced UHP metamorphism at the same peak P,T condition as its host eclogite. The very high Ca-Tschermakite content (31,34 mol.%) of the primary clinopyroxene indicates crystallization at about 9,17 kbar and 1250,1450 °C, and together with the microtextural observations, suggests that the protolith of the garnet pyroxenite was a cumulate from a former gabbroic body, in which case, the host eclogite might represent the gabbroic body. [source] INFLUENCE OF HYDROXYPROPYL METHYLCELLULOSE ON THE RHEOLOGICAL AND MICROSTRUCTURAL CHARACTERISTICS OF WHOLE WHEAT FLOUR DOUGH AND QUALITY OF PURIJOURNAL OF TEXTURE STUDIES, Issue 2 2009M. L. SUDHA ABSTRACT Puri is a traditional unleavened fried product prepared from whole wheat flour. Hydroxypropyl methylcellulose (HPMC) was used to study its effect on rheological characteristics of whole wheat flour dough and puri making quality. Addition of HPMC at 0.5 and 1.0% w/w increased the water absorption and dough stability whereas the resistance to extension and extensibility decreased. Pasting temperature, peak viscosity and cold paste viscosity gradually decreased. The moisture and fat contents of puri increased marginally. Quality parameters and sensory acceptability were monitored after 0 and 8 h of storage. Addition of 0.5% HPMC gave higher sensory scores. Microscopic observations during puri processing showed that the starch granules in the control dough were clearly visible in the protein matrix, which reduced on frying due to partial gelatinization. Microstructure of puri with HPMC showed higher gelatinization of starch. It also helped in moisture retention and hence, resulted in highly pliable and soft-textured puri. PRACTICAL APPLICATIONS Puri is a traditional unleavened fried product that is prepared by mixing whole wheat flour and water, sheeted to a desirable thickness and fried. Use of hydroxypropyl methylcellulose (HPMC) affected the whole wheat flour dough and puri making quality. It helped in moisture retention and hence, resulted in highly pliable and soft-textured puri. Microstructure of puri with HPMC showed higher gelatinization of starch. [source] Multiscale observation of biological interactions of nanocarriers: From nano to macroMICROSCOPY RESEARCH AND TECHNIQUE, Issue 9 2010Su-Eon Jin Abstract Microscopic observations have played a key role in recent advancements in nanotechnology-based biomedical sciences. In particular, multiscale observation is necessary to fully understand the nano-bio interfaces where a large amount of unprecedented phenomena have been reported. This review describes how to address the physicochemical and biological interactions of nanocarriers within the biological environments using microscopic tools. The imaging techniques are categorized based on the size scale of detection. For observation of the nanoscale biological interactions of nanocarriers, we discuss atomic force microscopy (AFM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). For the micro to macro-scale (in vitro and in vivo) observation, we focus on confocal laser scanning microscopy (CLSM) as well as in vivo imaging systems such as magnetic resonance imaging (MRI), superconducting quantum interference devices, and IVIS®. Additionally, recently developed combined techniques such as AFM-CLSM, correlative light and electron microscopy (CLEM), and SEM spectroscopy are also discussed. In this review, we describe how each technique helps elucidate certain physicochemical and biological activities of nanocarriers such as dendrimers, polymers, liposomes, and polymeric/inorganic nanoparticles, thus providing a toolbox for bioengineers, pharmaceutical scientists, biologists, and research clinicians. Microsc. Res. Tech. 73:813,823, 2010. © 2010 Wiley-Liss, Inc. [source] Pick's disease with Pick bodies combined with progressive supranuclear palsy without tuft-shaped astrocytes: A clinical, neuroradiologic and pathological study of an autopsied caseNEUROPATHOLOGY, Issue 3 2006Lu-Ning Wang We report clinical, neuroradiologic features, and neuropathologic findings of a 76-year-old man with coexistent Pick's disease and progressive supranuclear palsy. The patient presented with loss of recent memory, abnormal behavior and change in personality at the age of 60. The symptoms were progressive. Three years later, repetitive or compulsive behavior became prominent. About 9 years after onset, he had difficulty moving and became bed-ridden because of a fracture of his left leg. His condition gradually deteriorated and he developed mutism and became vegetative. The patient died from pneumonia 16 years after the onset of symptoms. Serial MRI scans showed progressive cortex atrophy, especially in the bilateral frontal and temporal lobes. Macroscopic inspection showed severe atrophy of the whole brain, including cerebrum, brainstem and cerebellum. Microscopic observations showed extensive superficial spongiosis and severe neuronal loss with gliosis in the second and third cortical layers in the frontal, temporal and parietal cortex. There were Pick cells and argyrophilic Pick bodies, which were tau- and ubiquitin-positive in neurons of layers II,III of the above-mentioned cortex. Numerous argyrophilic Pick bodies were observed in the hippocampus, especially in the dentate fascia. In addition, moderate to severe loss of neurons was found with gliosis and a lot of Gallyas/tau-positive globus neurofibrillary tangles in the caudate nucleus, globus pallidus, thalamus, substantia nigra, locus coeruleus and dentate nucleus. Numerous thorned-astrocytes and coiled bodies but no-tuft shaped astrocytes were noted in the basal ganglion, brainstem and cerebellar white matter. In conclusion, these histopathological features were compatible with classical Pick's disease and coexistence with progressive supranuclear palsy without tuft-shaped astrocytes. [source] Abscisic acid determines arbuscule development and functionality in the tomato arbuscular mycorrhizaNEW PHYTOLOGIST, Issue 3 2007María José Herrera-Medina Summary ,,The role of abscisic acid (ABA) during the establishment of the arbuscular mycorrhiza (AM) was studied using ABA sitiens tomato (Lycopersicon esculentum) mutants with reduced ABA concentrations. ,,Sitiens plants and wild-type (WT) plants were colonized by Glomus intraradices. Trypan blue and alkaline phosphatase histochemical staining procedures were used to determine both root colonization and fungal efficiency. Exogenous ABA and silver thiosulfate (STS) were applied to establish the role of ABA and putative antagonistic cross-talk between ABA and ethylene during AM formation, respectively. ,,Sitiens plants were less susceptible to the AM fungus than WT plants. Microscopic observations and arbuscule quantification showed differences in arbuscule morphology between WT and sitiens plants. Both ABA and STS increased susceptibility to the AM fungus in WT and sitiens plants. Fungal alkaline phosphate activity in sitiens mutants was completely restored by ABA application. ,,The results demonstrate that ABA contributes to the susceptibility of tomato to infection by AM fungi, and that it seems to play an important role in the development of the complete arbuscule and its functionality. Ethylene perception is crucial to AM regulation, and the impairment of mycorrhiza development in ABA-deficient plants is at least partly attributable to ethylene. [source] Epidermal transpiration, ultrastructural characteristics and net photosynthesis of white spruce somatic seedlings in response to in vitro acclimatizationPHYSIOLOGIA PLANTARUM, Issue 4 2003Mohammed S. Lamhamedi Mortality of transplanted somatic seedlings at the stage of acclimatization is often high and likely due to rapid change in environmental conditions. To investigate the potential of in vitro acclimatization of somatic seedlings before soil transfer, somatic seedlings of white spruce (Picea glauca[Moench] Voss) were germinated on a liquid medium supplemented with sucrose. After 6 weeks in germination, sucrose was omitted from the medium for a supplementary 6 weeks at which time somatic seedlings were acclimatized in vitro in their germination tubes before transfer to soil. In vitro acclimatization of somatic seedlings was realized by transferring the test tubes containing the germinated somatic seedlings to the greenhouse for 9 days. During this period, the culture tube lids of acclimatized somatic seedlings were lifted progressively increasing air exchange between the tube and the greenhouse whereas, for non-acclimatized somatic seedlings the culture tubes were maintained closed during in vitro acclimatization. In vitro acclimatized somatic seedlings had higher asymptotic net photosynthesis (Pn) at light saturation than non-acclimatized seedlings (6 versus 4.5 µmol m,2 s,1). At the end of the in vitro acclimatization period, a lower rate of epidermal transpiration was also observed for acclimatized somatic seedlings (3.85 versus 4.75% h,1). Microscopic observations showed that starch granules were more abundant in needles of acclimatized somatic seedlings than in non-acclimatized somatic seedlings, probably as a result of their greater photosynthetic capacity. Needles from acclimatized somatic seedlings also showed more epicuticular wax projections than needles from non-acclimatized somatic seedlings. These structural changes may help somatic seedlings to restrict epidermal water loss and stomatal aperture. [source] USE OF PORCINE SMALL INTESTINAL SUBMUCOSA IN BLADDER AUGMENTATION IN RABBIT: LONG-TERM HISTOLOGICAL OUTCOMEANZ JOURNAL OF SURGERY, Issue 1-2 2008Ali Ayyildiz Aim: To investigate long-term histological features of bladder augmentation using porcine small intestine submucosa (SIS) in a rabbit model. Materials and method: Sixteen New Zealand rabbits were used. Porcine SIS was provided by a manufactured formation derived from the pig. After partial cystectomy was carried out on the bladder, a single layer of SIS (Cook® -SIS Technology, Cook Biotech Incorporated, West Lafayette, IN, USA) (2 × 5 cm) was sewn to bladder with continuous 5/0 vicryl suture material in a watertight manner. Urinary diversion was not used. The rabbits were killed 12 months later and perivesical fat was removed together with bladder. The 5-,m preparations taken from the samples were stained with haematoxylin,eosin and Mason's trichrome dye. S-100 and F8 stains were also used for immunohistochemical investigations. Results: The macroscopic view of bladder was normal. SIS was indistinguishable from normal bladder wall, but the region of the graft had a slight white coloration. Microscopic observations showed the continuity of transitional epithelium of host bladder tissue on SIS material. Detrusor and serosal layers were formed and these layers were indistinguishable from host bladder. Fibroblasts were scattered among the collagen fibrils. New vessel formations were present without lymphatic proliferation. Nerve regeneration was excellent. No inflammation was observed in normal and regenerated bladder wall. Conclusion: At the end of 12 months, the long-term histological features of bladder augmentation with porcine SIS in a rabbit model, such as presence of new vessel formations, nerve regeneration, collagen and smooth muscle regenerations, which were indistinguishable from original bladder, and the absence of inflammation, showed that SIS seems to be a viable alternative to the use of intestine in bladder augmentation. [source] Calyculin-A, an inhibitor for protein phosphatases, induces cortical contraction in unfertilized sea urchin eggsCYTOSKELETON, Issue 4 2001Yukako Asano Abstract When an unfertilized sea urchin egg was exposed to calyculin-A (CL-A), an inhibitor of protein phosphatases, for a short period and then lysed, the cortex contracted to exclude cytoplasm and became a cup-shaped mass. We call the contracted cortex "actin cup" since actin filaments were major structural components. Electron microscopic observation revealed that the cup consisted of inner electron-dense layer, middle microfilamentous layer, and outermost granular region. Microfilaments were heavily accumulated in the inner electron-dense layer. The middle layer also contained numerous microfilaments, which were determined to be actin filaments by myosin S1 decoration, and they were aligned so that their barbed ends directed toward the outermost region. Myosin II, Arp2, Arp3, and spectrin were concentrated in the actin cup. Immuno-electron microscopy revealed that myosin II was localized to the electron-dense layer. We further found that the cortical tension of the egg increased just after application of CL-A and reached maximum within 10 min. Cytochalasin B or butanedione monoxime blocked the contraction, which suggested that both actin filaments and myosin ATPase activity were required for the contraction. Myosin regulatory light chain (MRLC) in the actin cup was shown to be phosphorylated at the activation sites Ser-19 and Thr-18, by immunoblotting with anti-phosphoepitope antibodies. The phosphorylation of MRLC was also confirmed by a 32P in vivo labeling experiment. The CL-A-induced cortical contraction may be a good model system for studying the mechanism of cytokinesis. Cell Motil. Cytoskeleton 48:245,261, 2001. © 2001 Wiley-Liss, Inc. [source] A new morphospecies of Microcystis sp. forming bloom in the Cheffia dam (Algeria): Seasonal variation of microcystin concentrations in raw water and their removal in a full-scale treatment plantENVIRONMENTAL TOXICOLOGY, Issue 4 2007Hichèm Nasri Abstract Toxic cyanobacterial blooms are an increasing problem in Algeria. The production of cyanotoxins (microcystins) and their presence in drinking water represent growing hazards to human health. In this study, seasonal variations in the concentrations of total microcystins and physicochemical parameters (pH, temperature, dissolved oxygen, nitrate, orthophosphate, and chlorophyll- a) were analyzed in the Cheffia dam (Algeria), mainly used to supply drinking water. The removal of cyanobacterial cells and microcystins was also evaluated in full-scale plant associated with the Cheffia reservoir. The levels of microcystins (MCYSTs) in both raw and drinking water were evaluated using the protein phosphatase type 2A (PP2A) inhibition test as MCYST-LR equivalents. Identification of microcystin variants was achieved by LC/MS/MS. During the period of study (March,December 2004), microscopic observation showed the dominance in the autumn months (September,November) of a new morphospecies of Microcystis sp. The MCYST-LR equivalent concentrations in raw water varied between 50.8 and 28,886 ng L,1. The highest level of toxins was observed in October 2004 and was significantly correlated with the chlorophyll- a. Three variants of microcystins assigned as microcystin-YR (MCYST-YR), microcystin-LR (MCYST-LR), and 6Z -Adda stereoisomer of MCYST-LR were observed in the crude extract of the Microcystis sp. bloom sample. During the bloom period, total elimination of Microcystis sp. and toxins were achieved through a classical treatment plant comprised of coagulation and flocculation, powdered activated carbon at 15 mg L,1, slow sand filtration and chlorination before storage. © 2007 Wiley Periodicals, Inc. Environ Toxicol 22: 347,356, 2007. [source] An animal model of testicular toxicity by cyclosporine: evaluation and protectionFUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 2 2009Ridha Ben Ali Abstract CyclosporineA (CsA) improves the survival of patients who benefited from transplantation. However, its use is generally limited by its side effects. The aim of our study was to measure, in an experimental model, the changes of the testosterone plasma levels after 21 days of CsA treatment and to explain the mechanism of this modification. After treatment, the levels of CsA, testosterone, corticosterone, transaminases were measured. The cytotoxic effect of CsA was evaluated by microscopic observation. The experimental study showed that CsA had no effect on the plasmatic levels of hepatic enzymes - alanine aminotransferase, aspartate aminotransferase and gamma-glutamyl-transferase , because their plasma concentrations in treated rats did not differ from those of the sham group. The plasma concentration of corticosterone was not modified, the plasma level of testosterone decreased when the dose of cyclosporine was increased to 4 mg/kg/day. The photonic microscope observation showed that the number of Leydig cells was increased and the electronic microscope observation showed mitochondria alteration. The treatment by CsA and trimetazidine did not correct the alteration caused by CsA. N-benzyl-N'-(2-hydrox-3, 4-dimethyloxybenzyl)-pipeazine did not protect the mitochondrial function but partially protected mitochondria structure from the deleterious effect induced by CsA. The decrease of the plasma level of testosterone induced by CsA was due to the inhibition of the mitochondrial 20,22 desmolase which blocked the formation of the testosterone precursor and the destruction of the mitochondria structure. [source] Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA libraryGENES TO CELLS, Issue 3 2000Da-Qiao Ding Background Intracellular localization is an important part of the characterization of a gene product. In an attempt to search for genes based on the intracellular localization of their products, we constructed a green fluorescent protein (GFP)-fusion genomic DNA library of S. pombe. Results We constructed the S. pombe GFP-fusion genomic DNA library by fusing, in all three reading frames, random fragments of genomic DNA to the 5, end of the GFP gene in such a way that expression of potential GFP-fusion proteins would be under the control of the own promoters contained in the genomic DNA fragments. Fission yeast cells were transformed with this plasmid library, and microscopic screening of 49 845 transformants yielded 6954 transformants which exhibited GFP fluorescence, of which 728 transformants showed fluorescence localized to distinct intracellular structures such as the nucleus, the nuclear membrane, and cytoskeletal structures. Plasmids were isolated from 516 of these transformants, and a determination of their DNA sequences identified 250 independent genes. The intracellular localizations of the 250 GFP-fusion constructs was categorized as an image database; using this database, DNA sequences can be searched for based on the localizations of their products. Conclusions A number of new intracellular structural components were found in this library. The library of GFP-fusion constructs also provides useful fluorescent markers for various intracellular structures and cellular activities, which can be readily used for microscopic observation in living cells. [source] Scanning electron microscopic observation of oil/wax/water/surfactant systemINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 2 2005K. Ikuta We observed the internal structure of an oil/wax/water/surfactant system using a scanning electron microscope to investigate the relationship between its hardness and state of wax crystal. The molten wax (hydrogenated jojoba oil, ceresin, polyethylene wax, carnauba wax, or microcrystalline wax) was cast in a home-made spiral mold of aluminum foil for preparing the test specimen for SEM observation. In hydrogenated jojoba oil a fine frame-like crystal structure, the so-called ,card-house structure,' was observed but not in other waxes. The mixture of hydrogenated jojoba oil and water showed a few small droplets deposited on the roundish wax frame-like structure. On the other hand, waxes other than hydrogenated jojoba oil did not change their internal structure when they were mixed with water. This result suggested that hydrogenated jojoba oil showed uniquely high affinity for water. In the system of oil, water, surfactant, and various kinds of waxes, their crystal structure, hardness, and the shape of dispersed water particles were remarkably changed with the combination of waxes. In the system with ceresin and carnauba wax, the hardness measured by a card-tension meter was high, and the internal crystal structure was fine and amorphous. The water particle in the ceresin and carnauba wax system had a smaller diameter than that in the system containing hydrogenated jojoba oil. The system containing hydrogenated jojoba oil showed a card house-like wax crystal structure without high hardness. It was considered that the wax crystal structure played an important role in providing hardness and in contributing to the water distribution in the oil/wax/water/surfactant system. [source] Bacillus megaterium spore germination is influenced by inoculum sizeJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2002M.L. Caipo Aims:,The effect of spore density on the germination (time-to-germination, percent germination) of Bacillus megaterium spores on tryptic soy agar was determined using direct microscopic observation. Methods and Results:,Inoculum size varied from approximately 103 to 108 cfu ml,1 in a medium where pH=7 and the sodium chloride concentration was 0·5% w/v. Inoculum size was measured by global inoculum size (the concentration of spores on a microscope slide) and local inoculum size (the number of spores observed in a given microscope field of observation). Both global and local inoculum sizes had a significant effect on time-to-germination (TTG), but only the global inoculum size influenced the percentage germination of the observed spores. Conclusions:,These results show that higher concentrations of Bacillus megaterium spores encourage more rapid germination and more spores to germinate, indicating that low spore populations do not behave similarly to high spore populations. Significance and Impact of the Study:,A likely explanation for the inoculum size-dependency of germination would be chemical signalling or quorum sensing between Bacillus spores. [source] Cytotoxic action mode of a novel porphyrin derivative isolated from harmful red tide dinoflagellate Heterocapsa circularisquamaJOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 3 2008Daekyung Kim Abstract Heterocapsa circularisquama is known to cause lethal effect on bivalves, but toxic effect on fish has not been reported yet. Recently, we have found that H. circularisquama has potent light-dependent hemolytic toxins. Based on the chemical structural analysis, one of the hemolytic toxins named H2-a was found to be a novel porphyrin derivative with similar structure to pyropheophorbide a methyl ester (PME), a well-known photoactive hemolytic agent (Miyazaki et al., Aquatic Toxicol. 2005;73:382--393). To clarify the cytotoxic action mode of H2-a, we examined the effects of H2-a on HeLa cells in comparison with PME. The cytotoxicities of both reagents were strictly light dependent, and no significant cytotoxic effects including cellular morphological changes were induced without light illumination. The dose response curves revealed that H2-a showed stronger cytotoxicity to HeLa cells than PME. Fluorescence microscopic observation suggested that H2-a tends to accumulate in the plasma membrane, whereas PME seems to distribute entire cytoplasm. Although PME induced typical apoptotic nuclear morphological changes and DNA fragmentation in HeLa cells, no such apoptosis-inducing ability of H2-a was observed. Among the radical scavengers, histidine significantly inhibited the cytotoxic activity of H2-a, suggesting the involvement of singlet oxygen in the cytotoxicity. These results suggest that the cytotoxic mechanism of H2-a is necrotic rather than apoptosis differing from PME, even though these are structurally quite similar to each other. The relatively high affinity of H2-a to the plasma membrane might result in the potent and quick cytotoxicity without induction of apoptotic signal transduction. © 2008 Wiley Periodicals, Inc. J Biochem Mol Toxicol 22:158,165, 2008; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20216 [source] Occurrence of calcite in Sanbagawa pelitic schists: implications for the formation of garnet, rutile, oligoclase, biotite and hornblendeJOURNAL OF METAMORPHIC GEOLOGY, Issue 2 2002A. Goto Abstract The frequency of occurrence of minerals in 1876 samples of Sanbagawa pelitic schist in central Shikoku is summarized on the basis of microscopic observation accompanied, in part, by use of an electron microprobe. All samples contain quartz, plagioclase, phengite, chlorite and graphite. More than 90% of samples contain clinozoisite, titanite and apatite. Garnet is present in 95% of samples from the garnet zone, and biotite is present in 64% of samples from the albite-biotite zone. Calcite is found in about 40% of samples of the pelitic schist collected from outcrop, but occurs in 95% of the pelitic schist from drill cores. Calcite was apparently ubiquitous in the pelitic schist during the Sanbagawa metamorphism, but must have been dissolved recently by the action of surface or ground water. The mineral assemblages of the Sanbagawa pelitic schist have to be analyzed in the system with excess calcite, quartz, albite (or oligoclase), clinozoisite, graphite and fluid that is composed mainly of H2O, CO2 and CH4. In the presence of calcite, reactions that produce garnet, rutile, oligoclase, biotite and hornblende, some of which define isograds of the metamorphic belt, should be written as mixed volatile equilibria that tend to take place at lower temperature than the dehydration reactions that have been proposed. The presence of calcite in pelitic schist suggests that fluid composition is a variable as important in determining mineral assemblages as pressure and temperature. Thus Ca-bearing phases must be taken into account to analyze the phase relations of calcite-bearing pelitic schist, even if CaO content of Sanbagawa pelitic schist is low. As calcite is a common phase, the mineral assemblages of the biotite zone pelitic schist may contravene the mineralogical phase rule and warrant further study. [source] Promoted random orientation of the phenyl substituent of phenylhydroquinone,terephthalic acid polyesters prepared with a diphenyl chlorophosphate/pyridine condensing agentJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 10 2001Fukuji Higashi Abstract The reaction of sterically hindered hydroxyl groups at the 2-position of methylhydroquinone and phenylhydroquinone (PhHQ) to form esters was largely promoted by their slow addition to benzoic acid activated by diphenyl chlorophosphate in pyridine. A modification of this reaction was applied to the preparation of thermotropic terephthalic acid/PhHQ and 2,5-dichloroterephthalic acid/PhHQ polymers with randomly oriented phenyl substituents, and the properties of the polymers were studied in terms of their transition temperatures, which were determined by differential scanning calorimetry and microscopic observation. The melting points were lowered by about 30,50 °C by the dropwise addition of PhHQ over 10,30 min. The molecular structures of the 2,5-dichloroterephthalic acid/PhHQ polymers were studied by 13C NMR. © 2001 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 39: 1726,1732, 2001 [source] Actions of Acute and Chronic Ethanol on Presynaptic TerminalsALCOHOLISM, Issue 2 2006Marisa Roberto This article presents the proceedings of a symposium entitled "The Tipsy Terminal: Presynaptic Effects of Ethanol" (held at the annual meeting of the Research Society on Alcoholism, in Santa Barbara, CA, June 27, 2005). The objective of this symposium was to focus on a cellular site of ethanol action underrepresented in the alcohol literature, but quickly becoming a "hot" topic. The chairs of the session were Marisa Roberto and George Robert Siggins. Our speakers were chosen on the basis of the diverse electrophysiological and other methods used to discern the effects of acute and chronic ethanol on presynaptic terminals and on the basis of significant insights that their data provide for understanding ethanol actions on neurons in general, as mechanisms underlying problematic behavioral effects of alcohol. The 5 presenters drew from their recent studies examining the effects of acute and chronic ethanol using a range of sophisticated methods from electrophysiological analysis of paired-pulse facilitation and spontaneous and miniature synaptic currents (Drs. Weiner, Valenzuela, Zhu, and Morrisett), to direct recording of ion channel activity and peptide release from acutely isolated synaptic terminals (Dr. Treistman), to direct microscopic observation of vesicular release (Dr. Morrisett). They showed that ethanol administration could both increase and decrease the probability of release of different transmitters from synaptic terminals. The effects of ethanol on synaptic terminals could often be correlated with important behavioral or developmental actions of alcohol. These and other novel findings suggest that future analyses of synaptic effects of ethanol should attempt to ascertain, in multiple brain regions, the role of presynaptic terminals, relevant presynaptic receptors and signal transduction linkages, exocytotic mechanisms, and their involvement in alcohol's behavioral actions. Such studies could lead to new treatment strategies for alcohol intoxication, alcohol abuse, and alcoholism. [source] | ||||||||||||||||||||||||||||||||||