Disabling VEGF-Response of Purkinje Cells by Downregulation of KDR via miRNA-204-5p

The vascular endothelial growth factor (VEGF) is well known for its wide-ranging functions, not only in the vascular system, but also in the central (CNS) and peripheral nervous system (PNS). To study the role of VEGF in neuronal protection, growth and maturation processes have recently attracted much interest. These effects are mainly mediated by VEGF receptor 2 (VEGFR-2). Current studies have shown the age-dependent expression of VEGFR-2 in Purkinje cells (PC), promoting dendritogenesis in neonatal, but not in mature stages. We hypothesize that microRNAs (miRNA/miR) might be involved in the regulation of VEGFR-2 expression during the development of PC. In preliminary studies, we performed a miRNA profiling and identified miR204-5p as a potential regulator of VEGFR-2 expression. In the recent study, organotypic slice cultures of rat cerebella (postnatal day (p) 1 and 9) were cultivated and VEGFR-2 expression in PC was verified via immunohistochemistry. Additionally, PC at age p9 and p30 were isolated from cryosections by laser microdissection (LMD) to analyse VEGFR-2 expression by quantitative RT-PCR. To investigate the influence of miR204-5p on VEGFR-2 levels in PC, synthetic constructs including short hairpin (sh)-miR204-5p cassettes (miRNA-mimics), were microinjected into PC. The effects were analysed by confocal laser scanning microscopy (CLSM) and morphometric analysis. For the first time, we could show that miR204-5p has a negative effect on VEGF sensitivity in juvenile PC, resulting in a significant decrease of dendritic growth compared to untreated juvenile PC. In mature PC, the overexpression of miR204-5p leads to a shrinkage of dendrites despite VEGF treatment. The results of this study illustrate, for the first time, which miR204-5p expression has the potential to play a key role in cerebellar development by inhibiting VEGFR-2 expression in PC.

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First instar larvae of endemic Australian Miltogramminae (Diptera: Sarcophagidae)

Scientific Reports ◽

10.1038/s41598-020-80139-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Krzysztof Szpila ◽

Kinga Walczak ◽

Nikolas P. Johnston ◽

Thomas Pape ◽

James F. Wallman

Keyword(s):

Laser Scanning ◽

Instar Larva ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Maxillary Palpus ◽

Larval Morphology ◽

First Instar ◽

Morphological Differences ◽

Thoracic And Abdominal ◽

First Time

AbstractThe first instar larva of a species of the Australian endemic genus Aenigmetopia Malloch is described for the first time, along with the first instar larvae of three other Australian species representing the genera Amobia Robineau-Desvoidy and Protomiltogramma Townsend. Larval morphology was analysed using a combination of light microscopy, confocal laser scanning microscopy and scanning electron microscopy. The following morphological structures are documented: pseudocephalon, antennal complex, maxillary palpus, facial mask, modifications of thoracic and abdominal segments, anal region, spiracular field, posterior spiracles and details of the cephaloskeleton. Substantial morphological differences are observed between the three genera, most notably in the labrum and mouthhooks of the cephaloskeleton, sensory organs of the pseudocephalon, spinulation, sculpture of the integument and form of the spiracular field. The first instar larval morphology of Aenigmetopia amissa Johnston, Wallman, Szpila & Pape corroborates the close phylogenetic affinity of Aenigmetopia Malloch with Metopia Meigen, inferred from recent molecular analysis. The larval morphology of Amobia auriceps (Baranov), Protomiltogramma cincta Townsend and Protomiltogramma plebeia Malloch is mostly congruent with the morphology of Palaearctic representatives of both genera.

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Infection of human CD34+ progenitor cells with Bartonella henselae results in intraerythrocytic presence of B henselae

Blood ◽

10.1182/blood-2004-12-4670 ◽

2005 ◽

Vol 106 (4) ◽

pp. 1215-1222 ◽

Cited By ~ 56

Author(s):

Tanja Mändle ◽

Hermann Einsele ◽

Martin Schaller ◽

Diana Neumann ◽

Wichard Vogel ◽

...

Keyword(s):

Progenitor Cells ◽

Laser Scanning ◽

Bartonella Henselae ◽

Bacterial Pathogen ◽

Erythroid Differentiation ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Facs Analysis ◽

Human Cd34 ◽

First Time

Abstract Although there is evidence that endothelial cells are important targets for human pathogenic Bartonella species, the primary niche of infection is unknown. Here we elucidated whether human CD34+ hematopoietic progenitor cells (HPCs) internalize B henselae and may serve as a potential niche of the pathogen. We showed that B henselae does not adhere to or invade human erythrocytes. In contrast, B henselae invades and persists in HPCs as shown by gentamicin protection assays, confocal laser scanning microscopy (CLSM), and electron microscopy (EM). Fluorescence-activated cell sorting (FACS) analysis of glycophorin A expression revealed that erythroid differentiation of HPCs was unaffected following infection with B henselae. The number of intracellular B henselae continuously increased over a 13-day period. When HPCs were infected with B henselae immediately after isolation, intracellular bacteria were subsequently detectable in differentiated erythroid cells on day 9 and day 13 after infection, as shown by CLSM, EM, and FACS analysis. Our data provide, for the first time, evidence that a bacterial pathogen is able to infect and persist in differentiating HPCs, and suggest that HPCs might serve as a potential primary niche in Bartonella infections.

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Physiological controls on seawater uptake and calcification in the benthic foraminifer <i>Ammonia tepida</i>

Biogeosciences ◽

10.5194/bg-6-2669-2009 ◽

2009 ◽

Vol 6 (11) ◽

pp. 2669-2675 ◽

Cited By ~ 47

Author(s):

L. J. de Nooijer ◽

G. Langer ◽

G. Nehrke ◽

J. Bijma

Keyword(s):

Laser Scanning ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Free Calcium ◽

Benthic Foraminifer ◽

Amorphous Precursor ◽

Intracellular Ca ◽

Ca Ions ◽

First Time ◽

Cellular Ca

Abstract. To analyze the relation between seawater uptake and calcification, we incubated juveniles of the benthic foraminifer Ammonia tepida with various fluorescent probes and visualised them afterwards with confocal laser scanning microscopy. Vesicle membranes, Ca ions and vacuole fluids were followed with various tracers and showed for the first time that endocytosis of seawater is part of the calcification process in Ammonia tepida. Data on the intracellular Ca ion cycling allowed for calculating a preliminary cellular Ca budget during foraminiferal calcification. This showed that the free calcium involved in the production of a new chamber cannot be sufficient and suggests that foraminifera may precipitate their calcite from an amorphous precursor.

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Microscopic and Spectroscopic Analyses of Chlorhexidine Tolerance in Delftia acidovorans Biofilms

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02984-14 ◽

2014 ◽

Vol 58 (10) ◽

pp. 5673-5686 ◽

Cited By ~ 11

Author(s):

Tara Rema ◽

John R. Lawrence ◽

James J. Dynes ◽

Adam P. Hitchcock ◽

Darren R. Korber

Keyword(s):

Laser Scanning ◽

Cell Types ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Content Type ◽

Flow Cells ◽

Biofilm Thickness ◽

Delftia Acidovorans ◽

Scanning Transmission ◽

Negative Effect

ABSTRACTThe physicochemical responses ofDelftia acidovoransbiofilms exposed to the commonly used antimicrobial chlorhexidine (CHX) were examined in this study. A CHX-sensitive mutant (MIC, 1.0 μg ml−1) was derived from a CHX-tolerant (MIC, 15.0 μg ml−1)D. acidovoransparent strain using transposon mutagenesis.D. acidovoransmutant (MT51) and wild-type (WT15) strain biofilms were cultivated in flow cells and then treated with CHX at sub-MIC and inhibitory concentrations and examined by confocal laser scanning microscopy (CLSM), scanning transmission X-ray microscopy (STXM), and infrared (IR) spectroscopy. Specific morphological, structural, and chemical compositional differences between the CHX-treated and -untreated biofilms of both strains were observed. Apart from architectural differences, CLSM revealed a negative effect of CHX on biofilm thickness in the CHX-sensitive MT51 biofilms relative to those of the WT15 strain. STXM analyses showed that the WT15 biofilms contained two morphochemical cell variants, whereas only one type was detected in the MT51 biofilms. The cells in the MT51 biofilms bioaccumulated CHX to a similar extent as one of the cell types found in the WT15 biofilms, whereas the other cell type in the WT15 biofilms did not bioaccumulate CHX. STXM and IR spectral analyses revealed that CHX-sensitive MT51 cells accumulated the highest levels of CHX. Pretreating biofilms with EDTA promoted the accumulation of CHX in all cells. Thus, it is suggested that a subpopulation of cells that do not accumulate CHX appear to be responsible for greater CHX resistance inD. acidovoransWT15 biofilm in conjunction with the possible involvement of bacterial membrane stability.

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Progress towards Sustainable Control of Xylella fastidiosa subsp. pauca in Olive Groves of Salento (Apulia, Italy)

Pathogens ◽

10.3390/pathogens10060668 ◽

2021 ◽

Vol 10 (6) ◽

pp. 668

Author(s):

Marco Scortichini ◽

Stefania Loreti ◽

Nicoletta Pucci ◽

Valeria Scala ◽

Giuseppe Tatulli ◽

...

Keyword(s):

Control Strategy ◽

Laser Scanning ◽

Vascular System ◽

Xylella Fastidiosa ◽

Low Cost ◽

Interdisciplinary Studies ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Olive Groves

Xylella fastidiosa subsp. pauca is the causal agent of “olive quick decline syndrome” in Salento (Apulia, Italy). On April 2015, we started interdisciplinary studies to provide a sustainable control strategy for this pathogen that threatens the multi-millennial olive agroecosystem of Salento. Confocal laser scanning microscopy and fluorescence quantification showed that a zinc-copper-citric acid biocomplex—Dentamet®—reached the olive xylem tissue either after the spraying of the canopy or injection into the trunk, demonstrating its effective systemicity. The biocomplex showed in vitro bactericidal activity towards all X. fastidiosa subspecies. A mid-term evaluation of the control strategy performed in some olive groves of Salento indicated that this biocomplex significantly reduced both the symptoms and X. f. subsp. pauca cell concentration within the leaves of the local cultivars Ogliarola salentina and Cellina di Nardò. The treated trees started again to yield. A 1H-NMR metabolomic approach revealed, upon the treatments, a consistent increase in malic acid and γ-aminobutyrate for Ogliarola salentina and Cellina di Nardò trees, respectively. A novel endotherapy technique allowed injection of Dentamet® at low pressure directly into the vascular system of the tree and is currently under study for the promotion of resprouting in severely attacked trees. There are currently more than 700 ha of olive groves in Salento where this strategy is being applied to control X. f. subsp. pauca. These results collectively demonstrate an efficient, simple, low-cost, and environmentally sustainable strategy to control this pathogen in Salento.

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Pollen heteromorphism in Schleichera Lour. (Sapindaceae), observed in surface soil samples from central India

10.35535/acpa-2021-0003 ◽

2021 ◽

Vol 61 (1) ◽

pp. 32-41

Author(s):

Md. Firoze Quamar ◽

Biswajeet Thakur ◽

Veeru Kant Singh ◽

Santosh Kumar Pandey

Keyword(s):

Laser Scanning ◽

Surface Soil ◽

Pollen Grains ◽

Central India ◽

Soil Samples ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Pollen Heteromorphism ◽

C Value ◽

First Time

Angiosperms display striking variation of pollen morphological features within and between populations of the same species, as well as within individual plants. We describe and illustrate variation of pollen aperture number, which is called pollen heteromorphism, in Schleichera Lour. (Sapindaceae) from surface soil samples collected from central India, based on combined observations from light microscopy (LM) and confocal laser scanning microscopy (CLSM). Tri-zono-parasyncolporoidate pollen grains are, in general, known to occur in Schleichera Lour., but occasional tetra-zono-parasyncolporoidate pollen is also recorded, for the first time, from Chhattisgarh State, central India. Changes in ploidy level (diploidy/polyploidy), chromosome number, the C-value of DNA, completion of meiosis, as well as environmental factors and/or pollination ecology could be driving the occurrence of pollen heteromorphism. The present study could provide insights into the phylogeny and systematics, and has implications for pollen preservation as well.

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Mycoplasma suis Invades Porcine Erythrocytes

Infection and Immunity ◽

10.1128/iai.00773-08 ◽

2008 ◽

Vol 77 (2) ◽

pp. 576-584 ◽

Cited By ~ 77

Author(s):

K. Groebel ◽

K. Hoelzle ◽

M. M. Wittenbrink ◽

U. Ziegler ◽

L. E. Hoelzle

Keyword(s):

Laser Scanning ◽

Fluorescent Labeling ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Bacterial Cells ◽

Mycoplasma Suis ◽

Transmission Electron ◽

Blood Smears ◽

Hemotrophic Mycoplasma ◽

First Time

ABSTRACT Mycoplasma suis belongs to the hemotrophic mycoplasma group and causes infectious anemia in pigs. According to the present state of knowledge, this organism adheres to the surface of erythrocytes but does not invade them. We found a novel M. suis isolate that caused severe anemia in pigs with a fatal disease course. Interestingly, only marginal numbers of the bacteria were visible on and between the erythrocytes in acridine orange-stained blood smears for acutely diseased pigs, whereas very high loads of M. suis were detected in the same blood samples by quantitative PCR. These findings indicated that M. suis is capable of invading erythrocytes. By use of fluorescent labeling of M. suis and examination by confocal laser scanning microscopy, as well as scanning and transmission electron microscopy, we proved that the localization of M. suis was intracellular. This organism invades erythrocytes in an endocytosis-like process and is initially surrounded by two membranes, and it was also found floating freely in the cytoplasm. In conclusion, we were able to prove for the first time that a member of the hemotrophic mycoplasma group is able to invade the erythrocytes of its host. Such colonization should protect the bacterial cells from the host's immune response and hamper antibiotic treatment. In addition, an intracellular life cycle may explain the chronic nature of hemotrophic mycoplasma infections and should serve as the foundation for novel strategies in hemotrophic mycoplasma research (e.g., treatment or prophylaxis).

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The mandibular gland inNasonia vitripennis(Hymenoptera: Pteromalidae)

10.1101/006569 ◽

2014 ◽

Cited By ~ 3

Author(s):

István Mikó ◽

Andrew Deans

Keyword(s):

Confocal Laser Scanning Microscopy ◽

Laser Scanning ◽

Mandibular Gland ◽

Laser Scanning Microscopy ◽

Confocal Laser Scanning ◽

Scanning Microscopy ◽

Confocal Laser ◽

Nasonia Vitripennis ◽

Courtship Behaviors ◽

First Time

The mandibular gland ofNasonia vitripennis(Hymenoptera: Pteromalidae) is visualized for the first time, using Confocal Laser Scanning Microscopy and dissection. The gland was previously hypothesized to exist, based on observations of the wasp's courtship behaviors, but its presence had never been confirmed.

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Supplementary description of five species from the genus Cecidophyopsis (Eriophyoidea: Eriophyidae: Cecidophyinae)

Systematic and Applied Acarology ◽

10.11158/saa.24.8.15 ◽

2019 ◽

Vol 24 (8) ◽

pp. 1555-1578

Author(s):

Slavica Marinković ◽

Philipp Chetverikov ◽

Tatjana Cvrković ◽

Biljana Vidović ◽

Radmila Petanović

Keyword(s):

Laser Scanning ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Taxus Baccata ◽

Mitochondrial Cytochrome Oxidase Subunit ◽

Ilex Aquifolium ◽

Internal Genitalia ◽

Corylus Avellana L ◽

First Time ◽

Mitochondrial Cytochrome Oxidase

Supplementary morphological descriptions of five Cecidophyopsis species collected in Europe (Serbia, Austria, Italy and Montenegro) are given: Cecidophyopsis vermiformis from Corylus avellana L. (Betulaceae), C. hendersoni from Yucca gigantea Lem. (Asparagaceae), C. verilicis from Ilex aquifolium L. (Aquifoliaceae), C. psilaspis from Taxus baccata L. (Taxaceae) and C. malpighianus from Laurus nobilis L. (Lauraceae). Males of C. vermiformis, C. verilicis, C. hendersoni and C. malpighianus, and immatures of C. hendersoni and C. verilicis, are described for the first time. C. verilicis is recorded for the first time in the fauna of Serbia and the European region. Female cuticle-lined internal genitalia of five Cecidophyopsis species are studied under confocal laser scanning microscopy. A several steps of oviposition in cecidophyines is proposed based on CLSM observations on their internal genitalia. Sequences of the barcoding region of the mitochondrial cytochrome oxidase subunit I (mtCOI) gene are given for the following species: C. hendersoni, C. verilicis, C. psilaspis and C. malpighianus.

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Progressive Loss of Lymphatic Vessels in Skin of Patients with Systemic Sclerosis

The Journal of Rheumatology ◽

10.3899/jrheum.100767 ◽

2010 ◽

Vol 38 (2) ◽

pp. 297-301 ◽

Cited By ~ 13

Author(s):

MIRKO MANETTI ◽

ANNA FRANCA MILIA ◽

SERENA GUIDUCCI ◽

ELOISA ROMANO ◽

MARCO MATUCCI-CERINIC ◽

...

Keyword(s):

Systemic Sclerosis ◽

Laser Scanning ◽

Vascular System ◽

Lymphatic Vessels ◽

Connective Tissue Disorder ◽

Laser Scanning Microscopy ◽

Papillary Dermis ◽

Confocal Laser ◽

Skin Involvement ◽

Reticular Dermis

Objective.Systemic sclerosis (SSc) is a connective tissue disorder characterized by microvascular and fibrotic changes in the skin and internal organs. The role of blood vessel dysfunction in the pathogenesis of SSc has been extensively investigated, but few studies have addressed the involvement of the lymphatic vascular system. Our aim was to evaluate dermal lymphatic vessels in patients with SSc according to different phases of skin involvement.Methods.Skin biopsies were obtained from the forearm of 25 SSc patients (10 early/15 late-stage disease) and 13 healthy controls. Skin sections were immunostained for podoplanin (D2-40), which is selectively expressed in lymphatic endothelial cells, and examined by confocal laser scanning microscopy. Lymphatic vessels were counted in the papillary and reticular dermis. Data were analyzed using Student’s t test.Results.The number of lymphatic vessels was significantly reduced in the papillary and reticular dermis of SSc patients compared with controls. In early SSc, lymphatic vessel counts were not different from controls in the papillary dermis, and showed a trend toward a reduction in the reticular dermis. In late SSc, a significant reduction in lymphatic vessels compared with controls was found in both the papillary and reticular dermis. The number of lymphatic vessels in the papillary dermis of late SSc was significantly lower than in early SSc.Conclusion.In SSc, lymphatic microangiopathy is linked to the progression of skin involvement. The progressive disappearance of lymphatic vessels may have a critical pathogenetic role in the progression of SSc from an early edematous phase to overt fibrosis.

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