Anatomy and histochemistry of stromatal anamorphs in the Sclerotiniaceae

1989 ◽  
Vol 67 (2) ◽  
pp. 371-393 ◽  
Author(s):  
Linda M. Kohn ◽  
Douglas J. Grenville
Keyword(s):  
Extracellular Matrix ◽  
Cell Walls ◽  
Protein Body ◽  
Histochemical Staining ◽  
Sclerotinia Homoeocarpa ◽  
Anatomical Features ◽  
Sclerotium Cepivorum ◽  
Storage Reserve ◽  
Large Deposits

As part of comparative studies of stromata in the Sclerotiniaceae, mature sclerotial and substratal stromata produced in vitro by 19 species, and 1 form-species, representing 13 genera and 1 form-genus, were examined using light microscopy and histochemical staining. Sclerotial-stromatal taxa were Sclerotinia sclerotiorum, S. trifoliorum, S. minor. Sclerotium cepivorum, Botrytis cinerea, B. porri, Dumontinia tuberose, Ciborinia erythronii, Myriosclerotinia dennisii, M. borealis, Monilinia fructicola, and Stromatinia gladioli. Substratal-stromatal taxa were Lambertella subrenispora, Lanzia luteovirescens, Rutstroemia sydowiana, Stromatinia gladioli, Ovulinia azaleae, Sclerotinia homoeocarpa, Scleromitrula shiraiana, and Ciboria acerina. Histochemical staining, particularly 0.05% toluidine blue O in benzoate buffer at pH 4.4, was found to be useful in demarcating the zones within stromata: rind, cortex, and medulla. All sclerotia contained extensive reserves of carbohydrates in thick cell walls and copious extracellular matrix, while protein bodies were usually the major cytoplasmic storage reserve. A group of saprophytic, substratal isolates had thin medullary cell walls and less extracellular matrix, and did not store protein but stored large deposits of lipid in cytoplasm. A group of phytopathogenic, substratal-stromatal isolates appeared to be transitional, with anatomical features and extensive cytoplasmic protein body reserves suggesting that they produce indeterminate sclerotial stromata rather than true substratal stromata.

scholarly journals Combined Staining Techniques for Demonstration of Staphylococcus aureus Biofilm in Routine Histopathology

2018 ◽  
Vol 3 (1) ◽  
pp. 27-36 ◽  
Author(s):  
Louise Kruse Jensen ◽  
Nicole Lind Henriksen ◽  
Thomas Bjarnsholt ◽  
Kasper Nørskov Kragh ◽  
Henrik Elvang Jensen
Keyword(s):  
Staphylococcus Aureus ◽  
Extracellular Matrix ◽  
Bone Tissue ◽  
Human Case ◽  
Histochemical Staining ◽  
Aureus Strain ◽  
Bacterial Cells ◽  
Routine Basis ◽  
Bacterial Aggregates

Abstract. Aim: Visualization of Staphylococcus aureus biofilm using histochemical staining and combined histochemistry (HC) and immunohistochemistry (IHC).Methods: The ability of S. aureus S54F9 to form biofilm was tested in vitro. Hereafter, infected bone tissue was collected from two different porcine models of osteomyelitis inoculated with S. aureus strain S54F9. The infection time was five and fifteen days, respectively. Twenty-five different histochemical staining protocols were tested in order to find the stains that could identify extracellular biofilm matrix. Protocols with an optimal visualization of biofilm extracellular matrix were combined with an immunohistochemical protocol based on a specific antibody against S. aureus. The combined protocols were applied to the tissue from the porcine models and to infected bone tissue from a child suffering from chronic staphylococcal osteomyelitis for more than a year.Results: S. aureus S54F9 showed an ability to form biofilm in vitro. Visualization of biofilm, i.e. bacterial cells and extracellular matrix in different colours, was seen when the immunohistochemical protocol was combined with Alcian Blue pH3, Luna and Methyl-pyronin green. The bacterial cells were red to light brown and the extracellular matrix either light blue, blue or orange depending on the histochemical stain. In the porcine models and the human case 10 and 90 percent, respectively, of the bacterial aggregates in a 100x magnification field displayed both the extracellular matrix and the bacterial cells simultaneously in two different colours.Conclusions: A combination of HC and IHC can be used to diagnose and characterise biofilm infections on a routine basis.

Ultrastructure of stromatal anamorphs in the Sclerotiniaceae

1989 ◽  
Vol 67 (2) ◽  
pp. 394-406 ◽  
Author(s):  
Linda M. Kohn ◽  
Douglas J. Grenville
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Walls ◽  
Monilinia Fructicola ◽  
Cellular Organization ◽  
Ultrastructural Studies ◽  
Sclerotium Cepivorum ◽  
Transmission Electron ◽  
Septal Pores

As part of comparative anatomical, histochemical, and ultrastructural studies of stromata in the Sclerotiniaceae, mature stromata produced in vitro by 11 species representing six genera and one form-genus were examined using transmission electron microscopy. Sclerotial-stromatal taxa were Sclerotinia sclerotiorum, S. trifoliorum, S. minor, Sclerotium cepivorum, Botrytis cinerea, B. porri, Monilinia fructicola, and Myriosclerotinia borealis. Substratal-stromatal taxa were Sclerotinia homoeo-carpa, Rutstroemia sydowiana, and Lambertella subrenispora. Three types of rind were observed: a living cellular rind, a dead cellular rind, and a stromatal rind. Sclerotial species were distinguished from stromatal species not only by the rind type, but also by the confluent extracellular matrix around cortical and medullary cell walls. Presence of lacunae in this matrix distinguished Sclerotinia spp. and M. borealis from Botrytis spp. and Monilinia fructicola. Rind, cortical, and medullary cells contained abundant storage vacuoles in most taxa. The distribution and proportion of organelles to storage vacuoles differed among taxa. Plugged septal pores with associated Woronin bodies were similar among the taxa where they were observed. Sclerotia of Sclerotium cepivorum, which has no known teleomorph, are ultrastructurally most like sclerotia of Sclerotinia or Botrytis anamorphs of Botryotinia species. Substratal stromata of S. homoeocarpa showed unusually complex cellular organization. Sclerotial stromata of M. fructicola contained unusual storage vacuoles with heterogeneous contents.

Neutrophil migration through in vitro interstitial matrix

1992 ◽  
Vol 50 (1) ◽  
pp. 894-895
Author(s):  
J. Roemer ◽  
S.R. Simon
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Cell Migration ◽  
Smooth Muscle Cells ◽  
Inflammatory Cell ◽  
Neutrophil Migration ◽  
Culture Conditions ◽  
Aortic Smooth Muscle ◽  
Specific Culture

We are developing an in vitro interstitial extracellular matrix (ECM) system for study of inflammatory cell migration. Falcon brand Cyclopore membrane inserts of various pore sizes are used as a support substrate for production of ECM by R22 rat aortic smooth muscle cells. Under specific culture conditions these cells produce a highly insoluble matrix consisting of typical interstitial ECM components, i.e.: types I and III collagen, elastin, proteoglycans and fibronectin.

Development of an in vitro Model for the Study of the Response of Equine Tendon Fibroblasts to Injury and Medication

1997 ◽  
Vol 10 (01) ◽  
pp. 6-11 ◽  
Author(s):  
R. F. Rosenbusch ◽  
L. C. Booth ◽  
L. A. Dahlgren
Keyword(s):  
Electron Microscopy ◽  
Extracellular Matrix ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Tendon Healing ◽  
Matrix Proteins ◽  
Isolated Cells ◽  
Superficial Digital Flexor Tendon ◽  
Vitro Model

SummaryEquine tendon fibroblasts were isolated from explants of superficial digital flexor tendon, subcultured and maintained in monolayers. The cells were characterized by light microscopy, electron microscopy and radiolabel studies for proteoglycan production. Two predominant cell morphologies were identified. The cells dedifferentiated toward a more spindle shape with repeated subcultures. Equine tendon fibroblasts were successfully cryopreserved and subsequently subcultured. The ability to produce proteoglycan was preserved.The isolated cells were identified as fibroblasts, based on their characteristic shape by light microscopy and ultrastructure and the active production of extracellular matrix proteins. Abundant rough endoplasmic reticulum and the production of extracellular matrix products demonstrated active protein production and export. Proteoglycans were measurable via liquid scintillation counting in both the cell-associated fraction and free in the supernatant. This model is currently being utilized to study the effects of polysulfated glycosaminoglycan on tendon healing. Future uses include studying the effects of other pharmaceuticals, such as hyaluronic acid, on tendon healing.A model was developed for in vitro investigations into tendon healing. Fibroblasts were isolated from equine superficial digital flexor tendons and maintained in monolayer culture. The tenocytes were characterized via light and electron microscopy. Proteoglycan production was measured, using radio-label techniques. The fibroblasts were cryopreserved and subsequently subcultured. The cells maintained their capacity for proteoglycan production, following repeated subculturing and cryopreservation.

Perturbation of Platelet Adhesion to Endothelial Cells by Plasminogen Activation In Vitro

1997 ◽  
Vol 78 (02) ◽  
pp. 934-938 ◽  
Author(s):  
Hsiun-ing Chen ◽  
Yueh-I Wu ◽  
Yu-Lun Hsieh ◽  
Guey-Yueh Shi ◽  
Meei-Jyh Jiang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Platelet Adhesion ◽  
Treatment Time ◽  
Shape Change ◽  
Umbilical Vein ◽  
Tissue Type ◽  
Apical Surface ◽  
Plasminogen Activation

SummaryTo investigate whether the endothelium-platelet interactions may be altered by plasminogen activation, cultured human umbilical vein endothelial cells (ECs) were treated with tissue-type plasminogen activator (t-PA) in the presence of plasminogen, and platelet adhesion to ECs was subsequently measured by using a tapered flow chamber. Our results demonstrated that platelets adhered more readily to t-PA treated EC monolayer than to the control monolayer at all shear stress levels tested. This phenomenon was treatment time-dependent and dose-dependent, and it could be blocked by adding plasmin inhibitors, such as e-amino caproic acid and aprotinin. Adherent platelets on t-PA treated EC monolayer underwent more severe shape change than those on the control monolayer. While the extracellular matrix directly treated with t-PA attracted less platelets than the control matrix did, platelet adhesion to the matrix that was produced by t-PA-treated ECs was unaltered. These data suggest that t-PA treatment on ECs compromised antiplatelet-adhesion capability on their apical surface without altering the reactivity of their extracellular matrix towards platelets.

Desmopressin (DDAVP) Enhances Platelet Adhesion to the Extracellular Matrix of Cultured Human Endothelial Cells through Increased Expression of Tissue Factor

1997 ◽  
Vol 77 (05) ◽  
pp. 0975-0980 ◽  
Author(s):  
Angel Gálvez ◽  
Goretti Gómez-Ortiz ◽  
Maribel Díaz-Ricart ◽  
Ginés Escolar ◽  
Rogelio González-Sarmiento ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Tissue Factor ◽  
Platelet Adhesion ◽  
Umbilical Vein ◽  
Procoagulant Activity ◽  
Experimental Conditions ◽  
Chromogenic Assay

SummaryThe effect of desmopressin (DDAVP) on thrombogenicity, expression of tissue factor and procoagulant activity (PCA) of extracellular matrix (ECM) generated by human umbilical vein endothelial cells cultures (HUVEC), was studied under different experimental conditions. HUVEC were incubated with DDAVP (1, 5 and 30 ng/ml) and then detached from their ECM. The reactivity towards platelets of this ECM was tested in a perfusion system. Coverslips covered with DD A VP-treated ECMs were inserted in a parallel-plate chamber and exposed to normal blood anticoagulated with low molecular weight heparin (Fragmin®, 20 U/ml). Perfusions were run for 5 min at a shear rate of 800 s1. Deposition of platelets on ECMs was significantly increased with respect to control ECMs when DDAVP was used at 5 and 30 ng/ml (p <0.05 and p <0.01 respectively). The increase in platelet deposition was prevented by incubation of ECMs with an antibody against human tissue factor prior to perfusion. Immunofluorescence studies positively detected tissue factor antigen on DDAVP derived ECMs. A chromogenic assay performed under standardized conditions revealed a statistically significant increase in the procoagulant activity of the ECMs produced by ECs incubated with 30 ng/ml DDAVP (p <0.01 vs. control samples). Northern blot analysis revealed increased levels of tissue factor mRNA in extracts from ECs exposed to DDAVP. Our data indicate that DDAVP in vitro enhances platelet adhesion to the ECMs through increased expression of tissue factor. A similar increase in the expression of tissue factor might contribute to the in vivo hemostatic effect of DDAVP.

Methods of experimental polyploidization and their use in apple breeding

2020 ◽  
Vol 62 ◽  
pp. 85-90
Author(s):  
L. V. Tashmatova ◽  
O. V. Matsneva ◽  
T. M. Khromova ◽  
V. V. Shakhov
Keyword(s):  
Exposure Time ◽  
Cell Walls ◽  
Prolonged Exposure ◽  
Ornamental Plants ◽  
Colchicine Treatment ◽  
Apple Trees ◽  
Open Ground ◽  
High Concentrations ◽  
Diploid Gametes

The article presents methods of experimental polyploidy of fruit, berry and ornamental plants. The purpose of this review is to highlight the problems and prospects of polyploidization of plants in the open ground and in vitro culture and the possibility of their application for apple trees. For the purpose of obtaining apple tetraploids as donors of diploid gametes, seed seedlings were treated with a solution of colchicine in concentrations of 0.1-0.4 % for 24 and 48 hours. Colchicine concentrations of 0.3 % and 0.4 % at 48 hours of treatment had a detrimental eff ect on their development. As a result, tetraploids and chimeras were obtained from seeds from free pollination of the varieties Orlik, Svezhest, Kandil Orlovsky, as well as from seeds obtained from crossing the varieties Svezhest×Bolotovskoe, Moskovskoe Оzherel’e×Imrus, Girlyanda×Venyaminovskoe. The optimal concentration of colchicine was 0.1 %. Methods of colchicine treatment have been studied: 1) adding to the nutrient medium, colchicine concentration: 0.01%, 0.02%, exposure time 24h-19 days; 2) applying amitotic solution to the growth point, colchicine concentration: 0.1 %, 0.2 %, exposure time 24h-7 days. To increase the penetration of colchicine through the cell walls, a 0.1 % dimexide solution was used. Studies have shown that high concentrations and prolonged exposure to colchicine reduce the viability of explants.

Antifungal Proteins from Plant Latex

2020 ◽  
Vol 21 (5) ◽  
pp. 497-506
Author(s):  
Mayck Silva Barbosa ◽  
Bruna da Silva Souza ◽  
Ana Clara Silva Sales ◽  
Jhoana D’arc Lopes de Sousa ◽  
Francisca Dayane Soares da Silva ◽  
...  
Keyword(s):  
Cell Walls ◽  
Defense Mechanisms ◽  
Hydrolytic Enzymes ◽  
Fungal Species ◽  
Biologically Active ◽  
Protein Classification ◽  
Fungal Cell ◽  
Antifungal Proteins ◽  
Plant Latex

Latex, a milky fluid found in several plants, is widely used for many purposes, and its proteins have been investigated by researchers. Many studies have shown that latex produced by some plant species is a natural source of biologically active compounds, and many of the hydrolytic enzymes are related to health benefits. Research on the characterization and industrial and pharmaceutical utility of latex has progressed in recent years. Latex proteins are associated with plants’ defense mechanisms, against attacks by fungi. In this respect, there are several biotechnological applications of antifungal proteins. Some findings reveal that antifungal proteins inhibit fungi by interrupting the synthesis of fungal cell walls or rupturing the membrane. Moreover, both phytopathogenic and clinical fungal strains are susceptible to latex proteins. The present review describes some important features of proteins isolated from plant latex which presented in vitro antifungal activities: protein classification, function, molecular weight, isoelectric point, as well as the fungal species that are inhibited by them. We also discuss their mechanisms of action.

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