scholarly journals ULTRASTRUCTURAL STUDIES ON THE PERMEABILITY OF THE MESOTHELIUM TO HORSERADISH PEROXIDASE

1968 ◽  
Vol 37 (1) ◽  
pp. 123-138 ◽  
Author(s):  
Ramzi S. Cotran ◽  
Morris J. Karnovsky
Keyword(s):  
Electron Microscope ◽  
Horseradish Peroxidase ◽  
Intraperitoneal Injection ◽  
Intercellular Junctions ◽  
Ultrastructural Studies ◽  
Intercellular Clefts ◽  
Peritoneal Mesothelium ◽  
Physiologic Data

Peritoneal mesothelium was exposed for 2–60 min to solutions of horseradish peroxidase by incubation in vitro, or after intraperitoneal injection in vivo. Peroxidase was localized, with the electron microscope in the intercellular clefts of the mesothelium, often along their entire lengths, in vesicles adjoining or contiguous with the clefts, and along the peritoneal and basal surfaces of the cell, and also in intracytoplasmic vacuoles. The intercellular junctions of peroxidase-treated mesothelium did not differ from those of controls: open and closed junctions were present in both groups. Intercellular localization was also obtained when the mesothelium was exposed to peroxidase during or after fixation. Although intracellular absorption of peroxidase and its incorporation into larger vacuoles were observed, there was no clearcut evidence of vesicular transport across the mesothelium in these experiments. These findings are consistent with physiologic data which postulate that mesothelial transport can be accounted for, at least in part, by passive diffusion through a system of pores, and they suggest that these pores are located in the intercellular clefts.

Probing the ultrastructure of the mitotic and meiotic spindle in eggs: An expeditious approach based on semi-thin (0.25 μm) serial sections

1983 ◽  
Vol 41 ◽  
pp. 552-555
Author(s):  
Conly L. Rieder ◽  
S. Bowser ◽  
R. Nowogrodzki ◽  
K. Ross ◽  
G. Sluder
Keyword(s):  
Small Sample Size ◽  
Small Sample ◽  
Meiotic Spindle ◽  
Serial Sections ◽  
Mitotic Apparatus ◽  
Thin Sections ◽  
Cell Cycles ◽  
Ultrastructural Studies

Eggs have long been a favorite material for studying the mechanism of karyokinesis in-vivo and in-vitro. They can be obtained in great numbers and, when fertilized, divide synchronously over many cell cycles. However, they are not considered to be a practical system for ultrastructural studies on the mitotic apparatus (MA) for several reasons, the most obvious of which is that sectioning them is a formidable task: over 1000 ultra-thin sections need to be cut from a single 80-100 μm diameter egg and of these sections only a small percentage will contain the area or structure of interest. Thus it is difficult and time consuming to obtain reliable ultrastructural data concerning the MA of eggs; and when it is obtained it is necessarily based on a small sample size.We have recently developed a procedure which will facilitate many studies concerned with the ultrastructure of the MA in eggs. It is based on the availability of biological HVEM's and on the observation that 0.25 μm thick serial sections can be screened at high resolution for content (after mounting on slot grids and staining with uranyl and lead) by phase contrast light microscopy (LM; Figs 1-2).

Ultrastructural studies on the interaction of haemophilus influenzae with human endothelial cells in vitro

1990 ◽  
Vol 48 (3) ◽  
pp. 636-637
Author(s):  
D.J.P. Ferguson ◽  
M. Virji ◽  
H. Kayhty ◽  
E.R. Moxon
Keyword(s):  
Endothelial Cells ◽  
Haemophilus Influenzae ◽  
Intercellular Junctions ◽  
Blood Stream ◽  
Ultrastructural Studies ◽  
Endothelial Barriers ◽  
Serotype B ◽  
Meningitis In Children ◽  
Respiratory Epithelial

Haemophilus influenzae is a human pathogen which causes meningitis in children. Systemic H. influenzae infection is largely confined to encapsulated serotype b organisms and is a major cause of meningitis in the U.K. and elsewhere. However, the pathogenesis of the disease is still poorly understood. Studies in the infant rat model, in which intranasal challenge results in bacteraemia, have shown that H. influenzae enters submucosal tissues and disseminates to the blood stream within minutes. The rapidity of these events suggests that H. influenzae penetrates both respiratory epithelial and endothelial barriers with great efficiency. It is not known whether the bacteria penetrate via the intercellular junctions, are translocated within the cells or carried across the cellular barrier in 'trojan horse' fashion within phagocytes. In the present studies, we have challenged cultured human umbilical cord_vein endothelial cells (HUVECs) with both capsulated (b+) and capsule-deficient (b-) isogenic variants of one strain of H. influenzae in order to investigate the interaction between the bacteria and HUVEC and the effect of the capsule.

A novel design of wound bandage using heparin-polyvinylpyrrolidone/TiO2 nanocomposite to improved antibacterial treatment and burn wound healing effect: In vitro and in vivo evaluation

2021 ◽  
Vol 11 (11) ◽  
pp. 1808-1818
Author(s):  
Xiuli Li ◽  
Jigang Wang ◽  
Xin Li ◽  
Xiaoqian Hou ◽  
Hao Wang ◽  
...  
Keyword(s):  
Electron Microscope ◽  
Dermal Fibroblast ◽  
Synergistic Effects ◽  
Bacterial Species ◽  
Human Dermal Fibroblast ◽  
Burn Wound ◽  
In Vivo Evaluation ◽  
Closure Rate

In our current study, porous heparin-polyvinylpyrrolidone/TiO2 nanocomposite (HpPVP/TiO2) bandage were prepared via the incorporation of TiO2 into HpPVP hydrogels for biomedical applications such as burn infection. The effect of the HpPVP hydrogels and the nanoparticles of TiO2 composition on the functional group and the surface properties of the as-fabricated bandages were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray diffractometry (XRD). The presence of TiO2 nanoparticles created the internal structure of the HpPVP hydrogel that aids in a homogeneous porous structure, as indicated by the scanning electron microscope (SEM). The size distribution of the TiO2 nanoparticles was measured using a transmission electron microscope (TEM). The studies on the mechanical properties of the HpPVP hydrogel indicate that the addition of TiO2 nanoparticles increases its strength. The prepared HpPVP/TiO2 nanocomposite dressing has excellent antimicrobial activity were tested against bacterial species (Staphylococcus aureus and Escherichia coli) and has good biocompatibility against human dermal fibroblast cells (HFFF2) for biological applications. In addition, in vivo evaluations in Kunming mice exposed that the as-fabricated HpPVP/TiO2 nanocomposite bandages increased the wound curing and facilitated accelerate skin cell construction along with collagen development. The synergistic effects of the HpPVP/TiO2 nanocomposite hydrogel dressing material, such as its excellent hydrophilic nature, good bactericidal activity, biocompatibility and wound closure rate through in vivo test makes it a suitable candidate for burn infections.

scholarly journals Targeting Cancer Cell Tight Junctions Enhances PLGA-Based Photothermal Sensitizers’ Performance In Vitro and In Vivo

Pharmaceutics ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 43
Author(s):  
Victoria O. Shipunova ◽  
Vera L. Kovalenko ◽  
Polina A. Kotelnikova ◽  
Anna S. Sogomonyan ◽  
Olga N. Shilova ◽  
...  
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
3D Culture ◽  
Intercellular Junctions ◽  
Cell Junctions ◽  
Multicellular Spheroids ◽  
Mesenchymal Transition ◽  
Non Invasive ◽  
Order Of Magnitude

The development of non-invasive photothermal therapy (PTT) methods utilizing nanoparticles as sensitizers is one of the most promising directions in modern oncology. Nanoparticles loaded with photothermal dyes are capable of delivering a sufficient amount of a therapeutic substance and releasing it with the desired kinetics in vivo. However, the effectiveness of oncotherapy methods, including PTT, is often limited due to poor penetration of sensitizers into the tumor, especially into solid tumors of epithelial origin characterized by tight cellular junctions. In this work, we synthesized 200 nm nanoparticles from the biocompatible copolymer of lactic and glycolic acid, PLGA, loaded with magnesium phthalocyanine, PLGA/Pht-Mg. The PLGA/Pht-Mg particles under the irradiation with NIR light (808 nm), heat the surrounding solution by 40 °C. The effectiveness of using such particles for cancer cells elimination was demonstrated in 2D culture in vitro and in our original 3D model with multicellular spheroids possessing tight cell contacts. It was shown that the mean inhibitory concentration of such nanoparticles upon light irradiation for 15 min worsens by more than an order of magnitude: IC50 increases from 3 µg/mL for 2D culture vs. 117 µg/mL for 3D culture. However, when using the JO-4 intercellular junction opener protein, which causes a short epithelial–mesenchymal transition and transiently opens intercellular junctions in epithelial cells, the efficiency of nanoparticles in 3D culture was comparable or even outperforming that for 2D (IC50 = 1.9 µg/mL with JO-4). Synergy in the co-administration of PTT nanosensitizers and JO-4 protein was found to retain in vivo using orthotopic tumors of BALB/c mice: we demonstrated that the efficiency in the delivery of such nanoparticles to the tumor is 2.5 times increased when PLGA/Pht-Mg nanoparticles are administered together with JO-4. Thus the targeting the tumor cell junctions can significantly increase the performance of PTT nanosensitizers.

An Electron-Microscope Study of the in vitro Transformation of Human Leucocytes

1967 ◽  
Vol 2 (3) ◽  
pp. 359-370
Author(s):  
J. A. CHAPMAN ◽  
M. W. ELVES ◽  
J. GOUGH
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Messenger Rna ◽  
Human Peripheral Blood ◽  
Limited Extent ◽  
Single Particles ◽  
Blast Cells ◽  
Protein Secreting

Electron-microscope studies of cultured small lymphocytes from human peripheral blood transforming into larger blastoid cells in the presence of phytohaemagglutinin (PHA) show that the transformed cell possesses the preliminary stages of development of a protein-synthesizing system. The transformed blastoid cell has abundant ribosomes, although, in contrast with in vivo protein-secreting cells, many of these occur as single particles with only a small proportion Linked in polysomal clusters. Endoplasmic reticulum membranes occur to a very limited extent and with a marked paucity of attached ribosomal particles; the few attached particles are usually located in groups. Some endoplasmic reticulum membranes revealed degenerative changes in otherwise normal cells. A moderately well-developed Golgi apparatus was a characteristic feature of the cells. Apart from the relatively low proportion of polysomes, in vitro PHA-transformed blastoid cells are identical in fine structure to in vivo blast cells (otherwise known as immunoblasts, haemocytoblasts, etc.) occurring in the immune response. It is suggested that messenger-RNA production in PHA-stimulated transformed cells may be reduced and that this could explain the limited number of polysomes and the restricted development of the endoplasmic reticulum.

Regulation of intestinal goblet cell secretion. III. Isolated intestinal epithelium

1984 ◽  
Vol 247 (6) ◽  
pp. G674-G681 ◽  
Author(s):  
T. E. Phillips ◽  
T. H. Phillips ◽  
M. R. Neutra
Keyword(s):  
Intestinal Epithelium ◽  
Direct Action ◽  
Intercellular Junctions ◽  
Goblet Cells ◽  
Mucus Secretion ◽  
Cell Secretion ◽  
Adult Rats ◽  
Compound Exocytosis

Cholinergic secretagogues evoke mucus secretion from goblet cells in the crypts of small and large intestinal mucosa in vivo and in organ culture. It was not known whether this response reflected a direct action on epithelial cell receptors or an indirect effect involving intermediate neurons of the enteric nervous system. To resolve this, carbachol was applied to isolated intestinal epithelium maintained in vitro. Intact sheets of epithelium, measuring 10–200 mm2, were isolated from the ileum and colon of adult rats following short intravascular perfusion with 30 mM EDTA. The isolated epithelia lacked a basal lamina and cytoplasmic blebs formed on the basal cell surfaces, but cell ultrastructure was normal and intercellular junctions were intact. Autoradiography revealed that both goblet and columnar cells continued to incorporate [3H]glucosamine into nascent secretory macromolecules for at least 45 min after isolation. When exposed to 20 microM carbachol for 5 min, crypt goblet cells discharged their stored mucin granules by compound exocytosis, whereas goblet cells in portions of the epithelium derived from villi or mucosal surfaces were unresponsive. We conclude that cholinergic secretagogues act directly on crypt epithelial cells to elicit mucus secretion.

REGULATION OF THE METABOLISM OF PITUITARY NUCLEIC ACIDS IN FEMALE RATS

1980 ◽  
Vol 85 (1) ◽  
pp. 1-8 ◽  
Author(s):  
J. BÍRÓ
Keyword(s):  
Nucleic Acids ◽  
Intraperitoneal Injection ◽  
Anterior Pituitary ◽  
Rna Metabolism ◽  
Female Rats ◽  
Biphasic Effect ◽  
Lh Rh ◽  
Lh Releasing Hormone

SUMMARY Ovariectomy caused an increase in the metabolism of pituitary nucleic acids. This effect was reversed in vivo by a biphasic action of oestradiol-17β which first facilitated RNA metabolism after 8 h and then inhibited it 16 h after intraperitoneal injection. To analyse the origin of this biphasic effect the roles of LH releasing hormone (LH-RH) and hysterectomy were examined. Incorporation of uridine into the RNA of the anterior pituitary gland of female rats was inhibited both in vivo and in vitro by LH-RH. Hysterectomy augmented the increase in the RNA metabolism caused by ovariectomy whereas steroid-free uterine extracts inhibited the increase significantly. We have concluded that extrapituitary factors may be involved in the effects of oestrogen on the metabolism of pituitary nucleic acids.

THE ENZYMIC FORMATION OF 6-(METHYLMERCAPTO)PURINE RIBONUCLEOSIDE 5′-PHOSPHATE

1966 ◽  
Vol 44 (2) ◽  
pp. 229-245 ◽  
Author(s):  
Ian C. Caldwell ◽  
J. Frank Henderson ◽  
A. R. P. Paterson
Keyword(s):  
Tumor Cells ◽  
Blood Cells ◽  
Intraperitoneal Injection ◽  
Ehrlich Ascites Carcinoma ◽  
Carcinoma Cells ◽  
Ehrlich Ascites ◽  
Mouse Tissues ◽  
Enzymatic Methods

6-(Methylmercapto)purine ribonucleoside (Me6MPR) is efficiently phosphorylated in mouse tissues and in Ehrlich ascites carcinoma cells in vivo; tumor cells in vitro and cell-free extracts of the tumor also phosphorylate this analogue ribonucleoside. The product of this reaction has been identified by chemical and enzymatic methods and by its chromatographic behaviour as Me6MPR 5′-phosphate. The evidence presented in this report indicates that no other major metabolites of Me6MPR are formed.The phosphorylation of Me6MPR by cell-free tumor extracts requires ATP and Mn2+ (or Mg2+), and evidence is presented that the reaction is probably mediated by adenosine kinase.Me-14C-6MPR is rapidly taken up by most mouse tissues following its intraperitoneal injection. Forty minutes after injection of the labeled drug, the highest levels of radioactivity were found in intestine, liver, blood cells, lung, and spleen, in descending order; virtually no radioactivity was found in brain tissue or in blood plasma.

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