Shear banding and yield stress in soft glassy materials.

Shear localization is a generic feature of flows in yield stress fluids and soft glassy materials but is incompletely understood. In the classical picture of yield stress fluids, shear banding happens because of a stress heterogeneity. Using recent developments in magnetic resonance imaging velocimetry, we show here for a colloidal gel that even in a homogeneous stress situation shear banding occurs, and that the width of the flowing band is uniquely determined by the macroscopically imposed shear rate rather than the stress. We present a simple physical model for flow of the gel showing that shear banding (localization) is a flow instability that is intrinsic to the material, and confirm the model predictions for our system using rheology and light scattering.

Ultracytochemical localization of glucose-6-phosphatase in Chang rat hepatoma in vivo and in vitro.

The standard lead precipitation method was used for ultracytochemical localization of glucose-6-phosphatase (G-6-Pase) in the in vivo and in vitro forms of the Chang rat hepatoma and in the normal adult rat liver. Reaction product was visualized as very fine particulate within the cisternae of the nuclear envelope and endoplasmic reticulum. Cytochemically, the amount of the G-6-Pase reaction product in both forms of the tumor cells was obviously less than that in the normal hepatocytes. Apparently, the enzyme was not completely deleted from the hepatoma cells. The results supported some biochemical data of certain other hepatomas. The successful ultracytochemical localization of G-6-Pase in cultured hepatoma cells has not been reported previously.

Cytochemical study of concanavalin A binding sites and their mobility in normal, cystic fibrosis, and SV40 transformed human fibroblasts in vitro.

Concanavalin A (Con A) binding sites and their mobility were studied by peroxidase (Po) and ferritin labeling techniques in normal and SV40 transformed human fibroblasts. Binding sites were visualized either as osmium black of 3'3-diaminobenzidine (DAB) reactions or as ferritin particles. DAB reaction products were localized at the external surface of the plasma membrane and in some multivesicular bodies of fixed cells. The labeling was continuous in normal and SV40 transformed human fibroblasts. When living cells were treated with Con A-Po at 4 degrees C and incubated at 37 degrees C, both normal and transformed cells showed remarkable changes. The foci of membrane indentations (caps or patches) are formed on the cell surface. Many labeled internalized vacuoles and vesicles appeared within the cytoplasm and in close proximity to the Golgi region of all cell types. The cellular changes occurred more quickly in transformed cells than in normal cells. It is concluded that normal cells do cap under certain conditions and that the plasma membranes of transformed cells are more fluid than those of normal cells.

The effects of ethylene thiourea administration upon rat liver cells.

The effect of ethylene thiourea (ETU) upon liver cells was evaluated in male Sprague-Dawley rats. ETU was administered ad libitum in drinking water at concentrations of 1, 5, 50, and 500 ppm for time intervals of 1, 2, and 5 days, 1 and 2 weeks, 1, 2, 4, and 8 months. Two additional groups of control animals received ETU-free drinking water or a diet supplemented with 0.06% 3-MeDAB. Electron microscopic evaluation of tissue samples could detect no changes in liver cell morphology of rats receiving 1, 5, or 50 ppm ETU for up to 8 months. By contrast, rats receiving 500 ppm ETU exhibited alterations in hepatic cell morphology after 4 months of exposure. These alterations included a dramatic increase in the amount of smooth endoplasmic reticulum (SER) with a concomitant reduction in rough endoplasmic reticulum (RER), and a relocation of microbodies and mitochondria to the periphery of the SER. No alterations were seen at the shorter time intervals. These changes probably represent a response to the sustained ingestion of high concentrations of highly toxic materials and most likely do not represent a specific response to ETU. No tumors were detected in any of the samples examined or in controls receiving ETU-free drinking water. Animals receiving 3-MeDAB in their diet all developed hepatic tumors within 4 months.

The distribution of cationized ferritin receptors on ciliated epithelial cells of rat trachea.

The interaction of tracheal cilia with the biphasic mucus layer covering the surface of the mammalian respiratory tract may be influenced by many cell surface coat components including those having an overall negative charge. In order to assess the distribution of ciliary anionic sites, cationized ferritin (CF) was used to label the surface of rat tracheal epithelium. If pieces of trachea were fixed with 3% glutaraldehyde and treated with CF at low (L) (0.08 mg/ml), medium (M) (0.32 mg/ml PBS), or high (H) (0.64 mg/ml PBS) concentrations, the label was distributed evenly over the entire external surface of the ciliary membrane at all concentrations. Unfixed tracheal tissue was also treated with L, M, and H CF for 1 or 5 min at 4 degrees C in order to minimize lateral redistribution of CF receptors. To ensure accessibility of the cell surface to CF the samples were agitated thoroughly during exposure. Exposure for 1 min to L, M, and H CF resulted in a light binding of ferritin particles on all portions of the ciliary membrane with occasional areas of multilayered binding distributed randomly on the ciliary shaft. When unfixed trachea was treated with CF for 5 min at 4 degrees C, CF binding was similar except heavier and more uniform. In no instance was there any preferential binding of CF to the ciliary tips at any of the concentrations used. Moreover, as indicated by the CF binding pattern at L concentrations, high density negative charges are present over almost the entire surface of the cilium. These results suggest that, unlike the ciliary membrane of other organs such as oviduct, negatively charged cell surface coat molecules are present on all areas of the ciliary membrane of rat tracheal epithelia.

Immunocytochemical localization of tubulin with colloidal gold in cilia of rabbit tracheal epithelial cultures.

Ciliated tracheal epithelia cell cultures were investigated immunocytochemically with anti-tubulin and colloidal gold. When rabbit tracheal cultures were fixed in paraformaldehyde, treated with acetone, anti-tubulin and a second antibody coupled to FITC, fluorescence was associated with cytoskeletal and axonemal microtubules. Cilia covering the apical surface of the ciliated tracheal cells fluoresced very brightly thus facilitating identification of this cell type. Electron microscopy of tracheal cultures fixed as above, treated with Triton-X 100 and incubated in anti-tubulin and protein A coupled to colloidal gold resulted in the highly specific localization of tubulin in ciliary axonemes and basal bodies. Omission of primary or secondary antibody resulted in extremely low levels of fluorescence while no colloidal gold particles could be detected in cultures at the electron microscopy level when rabbit anti-tubulin was omitted.

The development of ciliated and mucus cells from basal cells in hamster tracheal epithelial cell cultures.

Hamster tracheal epithelia consist of three cell types: ciliated, mucus and basal cells. Autoradiographic data from several studies suggest that either basal or non-ciliated columnar cells may serve as stem cells for regeneration of lost or damaged ciliated and mucus cells. The objective of the present study was to examine the role of basal cells in the formation of ciliated and mucus cells in hamster tracheal epithelial (HTE) cell cultures via tritiated thymidine ([3H]-TdR) autoradiography. When 3 day cultures were pulsed with [3H]-TdR for 6 hr and incubated for 2 additional days in non-radioactive media (5 day total) label was present in the nuclei of basal and columnar epithelial cells suggesting that the labeled columnar cells may be derived from basal cells. However, the morphological reorganization occurring during this 2 day interval may create difficulties in this interpretation. Since these morphological changes are minimal during the 6 day to 8 day in vitro period, 6 day HTE cultures were pulsed with [3H]-TdR for 6 hr and incubated for 2 additional days in non-radioactive media (8 day total), and examined to further study the fate of labeled basal cells during this period. Analysis of these 8 day cultures revealed that labeled nuclei were present in both basal cells and adjacent ciliated and mucus cells. These results do not exclude the possibility of non-basal cell origin of ciliated and mucus cells in other systems but suggest that, at least in HTE cultures, undifferentiated basal cells have the ability to develop into ciliated and mucus cells.

Organization of cytokeratin intermediate filaments in basal cells of growing rat trachea.

The basal cell in airway epithelium plays a major role in attachment of ciliated and nonciliated columnar cells to the basal lamina. As the airway grows in diameter and the columnar epithelium in height, the number of basal cells and the amount of tonofilaments (cytokeratin filaments) and anchoring junctions increase. In this way they maintain a constant attachment strength between the increased volume of the epithelium and the basal lamina. The purpose of this study was to determine which cytokeratins (CKs) are expressed in growing basal cells of the rat and demonstrate where they are localized in the cytoskeleton. Sprague Dawley rats 10, 30 and 90 days of age were used in this study. For light microscopy, tracheal samples were fixed in 95% alcohol or 4% formalin for 2 hr and then embedded in paraffin. For electron microscopy, the tracheal samples were placed in 20 mM EDTA in HBSS media minus Ca++ and Mg++ at pH 7.4 for 60 min to permeabilize the cells and expose the intracellular structures. Antibodies to cytokeratins 7, 8, 10, 13 and 18 did not react to basal cells at any age studied. Antibodies to CKs 5 + 8, 14, 16 + 13, and 19 gave a positive reaction with basal cells at each age studied. Immunogold particles representing antibodies to CK 14 were heavily distributed over intermediate filaments making up the cytoskeleton. Both CK 16 + 13 and 19 were also over intermediate filaments but at a much lower density.(ABSTRACT TRUNCATED AT 250 WORDS)

In situ preparation of rat tracheal basal cells.

The primary function of basal cells is to attach columnar epithelium to the basal lamina. They may also have limited stem cell potential, but very little is known of other biological functions. Basal cells lie on the basal lamina beneath the ciliated and secretory cells and do not reach the surface of the epithelium. The position of the cell beneath the ciliated and secretory cell epithelium makes their in situ study difficult. In order to further aid in the study of basal cells, we have developed an in situ preparation technique in which ciliated and secretory cells are removed. Treatment of rat tracheas with a 20 mM Na2 EDTA solution, pH 7.4, results in partial removal of columnar epithelium from the basal lamina. The percent of denuded columnar epithelial cells per mm of basal lamina is 43.9 +/- 7.8% at 60 min, 47.6 +/- 8.4% at 90 min, and 52.6 +/- 2.7% at 120 min. The viability of the exposed basal cells was the same at both 60 and 90 min of treatment (79.4 +/- 7.8 and 78.0 +/- 8.5, respectively). Morphologically, the exposed basal cells are attached to the basal lamina by hemidesmosomes and are similar to those in the intact animal.

An in vitro system for the study of tracheal epithelial cells.

Enzymatically dissociated hamster tracheal epithelial (HTE) cells were cultured on collagen coated Millicell filters. Within 3-5 days after being placed in culture large numbers of ciliated and mucus cells began to appear. By 1 week the HTE cells closely resembled those seen in vivo, i.e. columnar morphology and organelle polarity. After 4 weeks in vitro the HTE cultures were still able to maintain the polarity and overall columnar morphology found in in vivo tissue. There was, in addition, no apparent degradation of the collagen substrate. Auto-radiographic data indicated that there was an initial period of high DNA synthesis during the first 3 days in vitro. This was followed by a second phase in which, by day 6, the amount of DNA synthesis was greatly reduced. Analysis of the numbers of ciliated cells relative to non-ciliated cells demonstrated that between days 5 and 8 there was an increase in the percentage of ciliated cells, suggesting that cellular differentiation (i.e. ciliogenesis) follows cellular proliferation. The results of this study show that when HTE cells are grown on collagen-coated Millicell filters there is a significant improvement in cell growth and morphology yielding cells that are very similar to those present under in vivo conditions. Moreover, since there is no degradation of the collagen substrate, HTE cultures may be suitable for long-term studies of respiratory tract epithelia.

An ultrastructural study of morphogenesis of fibrogranular complex and centriole in ductuli efferentes of Chinese hamster.

An ultrastructural study of ciliated epithelial cells in the ductuli efferentes of young and adult hamsters has revealed that these cells possess dense granules, dense granule clusters, dense bodies and fibrogranular complexes as reservoirs or precursors for ciliogenesis. The dense granules are first seen in the centrosomal region. Later, many dense granules and dense granule clusters appear in the apical portion of the epithelial cells where, subsequently, dense bodies are also found. Finally, the fibrogranular complexes are formed in adults. Morphological evidence strongly suggests that cilia are formed from diplosomal centrioles, de novo centrioles, dense body centrioles, and fibrogranular complex centrioles. Ciliogenesis begins in the fourth day after birth and increases rapidly in the fifth day. After the sixth day, cilia appear to be generated mostly from dense bodies and the total ciliogenesis activities gradually decrease as the animal ages.

The effect of anti-exotoxin A on the adherence of Pseudomonas aeruginosa to hamster tracheal epithelial cells in vitro.

One of the most important initial events of colonization and infection of epithelial tissues is the adherence of bacteria to mucosal surfaces. Bacterial adhesion to the epithelial cell may be mediated by a variety of adhesins, including exoproducts. One of these exoproducts, exotoxin A (EA) is a three-domain bacterial toxin that kills mammalian cells by gaining entry to the cytosol and inactivating protein synthesis. In the present study, HTE cultures, 2-4 weeks in vitro (containing both ciliated and non-ciliated cells), were treated for 1 hr with two different non-mucoid strains of Pseudomonas aeruginosa (1 x 10(8) organisms/ml) in the presence of anti-EA. 50 randomly selected fields were evaluated via SEM at x2500 magnification and the number of bacterial clusters/field quantitated. The results of this study indicate, first, that both piliated (ATCC15692) and non-piliated (PAKp) P. aeruginosa will bind to the HTE cells and, second, that treatment of HTE cells with either strain of P. aeruginosa in the presence of anti-EA will reduce bacterial binding by 25% to 50%. Thus, EA may participate in the adhesion of P. aeruginosa to respiratory tract epithelia.

Immunocytochemical localization of laminin in hamster tracheal epithelial cell cultures.

EM examination of 28 day cultures of enzymatically dissociated hamster tracheal epithelial (HTE) cells grown on collagen coated millipore filters reveals that fragments of basal lamina may be present at the basal plasmalemma. Since the basal lamina consists of several major components including type IV collagen, heparan sulfate proteoglycans, entactin/nidogen, and laminin, questions naturally arise concerning the presence of such a structure in this cell culture system. When immunocytochemical procedures utilizing anti-laminin antibody and PAP techniques are carried out with paraffin sections of HTE culture at 1,2,3, and 4 weeks in vitro, LM analysis reveals that a thin, dense line of reaction product is present between the basal surface of the HTE cells and the underlying collagen substrate. Immunoblotting evaluation carried out with supernatants of 7d HTE cell homogenates and HTE cell conditioned media also indicate that laminin is being produced by the tracheal cells. Thus, the presence of basal lamina-like fragments, the immunocytochemical localization of laminin, and immunoblot identification of laminin in hamster tracheal epithelial cell cultures, suggest that, although basal lamina components may be produced by HTE cells, at the time points tested, they are not yet being organized into a complete basal lamina.

Type IV collagen localization in hamster tracheal epithelial cell cultures.

Hamster tracheal epithelial (HTE) cell cultures when grown on collagen coated Millicell-HA filters for 28 d in vitro have an incomplete basal lamina-like structure at the basal plasmalemma/collagen interface. EM evaluation of 42 d HTE cell cultures also revealed the presence of hemidesmosome-like structures on the basal plasmalemma. In order to better characterize this discontinuous basal lamina-like material, HTE cultures at 7, 14, 21, and 28 d were evaluated for the presence of type IV collagen, a basal lamina component. Immunocytochemical treatment of HTE cultures at these time points resulted in the presence of reaction product at the base of the cell layer. When immunocytochemical procedures for the localization of type IV collagen were carried out with normal mouse trachea, the results were also positive. Immunoblotting evaluation of HTE cell supernatants and conditioned media also indicated the presence of type IV collagen. Taken altogether, the presence of what appears to be basal lamina, hemidesmosome-like structures and immunocytochemical and immunoblot data showing the presence of type IV collagen in HTE cultures suggest that HTE cells may be producing basal lamina components but cannot organize them into a complete basal lamina.

[Transurethral treatment of seminal vesicle abscess]. / Transuretral behandling af vesicula seminalis-absces.

An 83-year old male with a seminal vesicle abscess is presented. Initially, cancer of the prostate was suspected and the findings of the transrectal ultrasound scan were misinterpreted. The diagnosis was made by CT-scan. When drainage failed the patient was treated successfully with transurethral unroofing.

[Transurethral microwave thermotherapy. A therapeutic option for high-risk patients with urinary retention secondary to benign prostatic hyperplasia]. / Transuretral mikrobølgetermoterapi. En behandlingsmulighed hos højrisikopatienter med urinretention sekundaert til benign prostatahyperplasi.

Urinary retention secondary to benign prostatic hyperplasia is considered an absolute indication for surgical treatment of the prostate. Transurethral resection of the prostate is still considered the gold standard in terms of effectiveness. One of several new techniques for treatment of BPH is transurethral microwave thermotherapy (TUMT). Our first experiences with this technique in a group of patients with urinary retention were analysed retrospectively. In the short term 16 of the 25 treated patients were able to void spontaneously with acceptable bladder emptying (64%, 95% CI: 43-85). Later on, three of these initial successes received further treatments for BPH, two had a transurethral resection of the prostate and one was given medical therapy. No serious complication was seen except in one patient who developed a urethrorectal fistula which healed following conservative treatment. The success rate following TUMT was inferior to that of standard transurethral resection, but TUMT seems an acceptable alternative in patients with pronounced co-morbidity.

Ureterorenoscopy. Results of diagnostic and therapeutic procedures.

During a 32-month period, 215 ureterorenoscopies were attempted in 171 ureterorenal units. The success rates for stone retrieval were 87% in the lower third of the ureter, 67% in the middle third, 75% in the upper third, and 55% in the pelvis. In 18% of cases a particular lesion could not be reached in diagnostic endoscopy but biopsies were obtained from seven out of eight tumours. The ureteric orifice was never dilated beyond 8 F and, yet, only four times could the instrument not be passed into the ureter. One major complication, an avulsion of 12 cm of the distal ureter, occurred. It is concluded that ureterorenoscopy is of significant value in the diagnosis and treatment of ureteric diseases.

Ureteropyeloscopic tumour treatment.

Ureteropyeloscopy was used to follow-up two patients with upper urinary tract transitional cell tumours treated by topical chemotherapy. Epodyl was instilled into the pelvis and ureter through a ureteric catheter. There were no complications. The ureteric tumours responded completely, whereas a pelvic papilloma diminished in size to about 1 mm.