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.

Transurethral interstitial laser coagulation of the prostate and transurethral microwave thermotherapy vs transurethral resection or incision of the prostate: results of a randomized, controlled study in patients with symptomatic benign prostatic hyperplasia.

OBJECTIVE: To compare the efficacy and frequency of complications of transurethral interstitial laser coagulation (ILC) and transurethral microwave thermotherapy (TUMT) with transurethral resection or incision of the prostate (TURP/TUIP) in patients with symptomatic benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: Forty-eight patients were randomized to undergo ILC, 46 to TUMT and 24 to TURP/TUIP; they were followed for 6 months and the outcome analysed on an intention-to-treat basis. RESULTS: At 6 months the symptom scores and maximum urinary flow rate (Qmax) had improved significantly in all groups. At 6 months the mean symptom score was 9.2 in both experimental groups and 6.8 in the control group (P > 0.05); the mean Qmax was 20.6 mL/s in the control group, 16.2 in the ILC group (P > 0.05 vs control) and 13.2 in the TUMT group (P < 0.05 vs. the control group). In the TUMT group patients developing urinary retention afterward had a significantly greater increase in Qmax than those who did not. The types of complications in the three groups varied. Urinary tract infection occurred frequently in the experimental groups, especially after ILC, whereas the 'well-known' complications of TURP occurred in the control group. Overall, 36% in the ILC, 54% in the TUMT and 73% in the control group had no complications (retrograde ejaculation excluded) during the first 6 months. One patient in the TUMT group underwent TURP after 3 months, whereas no patients in the ILC or the con-trol group were re-treated for BPH within the first 6 months. CONCLUSION: In the short term both ILC and TUMT are reasonable alternatives to standard transurethral surgery for symptomatic BPH, where the reduction of symptoms is the primary goal of treatment. However, both ILC and TUMT were associated with morbidity, although the complication profiles differed from those after TURP/TUIP. Both ILC and TUMT seem advantageous in some patients because of the reduced risk of bleeding and the eliminated risk of TUR syndrome, and because TUMT only requires local anaesthesia. Thus, as neither treatment is better in all aspects, the advantages of one technique over the other must be weighed when deciding how to treat each patient.

Cost-effectiveness of new treatments for benign prostatic hyperplasia: results of a randomized trial comparing the short-term cost-effectiveness of transurethral interstitial laser coagulation of the prostate, transurethral microwave thermotherapy and standard transurethral resection or incision of the prostate.

OBJECTIVES: To compare the short-term cost-effectiveness of ILC and TUMT with that of transurethral resection or incision of the prostate in patients with symptomatic benign prostatic hyperplasia. MATERIAL AND METHODS: One hundred and eighteen patients were randomized to ILC, TUMT and TUR-P/TUI-P in 2:2:1 fashion. The use of resources within the first 6 months of follow-up were measured and the cost of treatment for each patient were calculated. A few parameters, primarily related to the time spent by the staff, were measured only in a subgroup of patients. A cost-effectiveness analysis was performed based on the average calculated cost and change in I-PSS found in each group. For costly resources a sensitivity analysis was performed. RESULTS: At 6 months the cost was lowest in the TUMT group and highest in the ILC group. The cost-effectiveness ratio (C/E) was 763 DKK/point reduction in I-PSS in the control group and 1.200 and 938 in the ILC and TUMT group, respectively. An incremental analysis demonstrated that TUR-P/TUI-P dominated ILC. In a similar comparison of the TUMT and the TUR-P/TUI-P group an incremental ratio of 170 DKK/extra point of reduction in I-PSS was found in the TUR-P/TUI-P group. CONCLUSION: In the short-term TUMT and TUR-P has comparable cost-effectiveness. TUR-P was slightly more effective than TUMT, but the cost was also slightly higher. In our set-up of ILC the short-term cost-effectiveness of ILC was inferior to that of TUR-P. Conclusions should be made with caution, since the follow-up at present is short.

[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.

[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.

Characteristics of 25-hydroxycholesterol-induced apoptosis in the human leukemic cell line CEM.

Cholesterol and related compounds can give rise to oxygenated sterol molecules (oxysterols) which are potent regulators of lymphoid cell growth. Oxysterols added exogenously cause cell death of several lines of cultured cells, and on the basis of limited criteria, it has been suggested that this death is apoptosis. In the present study, we show definitive evidence that 25-hydroxycholesterol (25OHC) kills cells of the clone CEM-C7 by apoptosis and establish the temporal sequence of related cellular and biochemical events. Cell shrinkage was evident as early as 12 h, while cell death was not evident until after 24 h. It mounted rapidly after that, and by 72 h, virtually all cells were dead. Electron microscopic analysis shows that by 24 h after treatment and before the onset of cell death, early ultrastructural features typical of apoptosis were present. DNA breaks were detected by TUNEL assay prior to the onset of cell death. Two types of specific DNA pieces often associated with apoptosis were found as increasing numbers of cells died. DNA fragments of 300 and 50 kbp were not appreciable until 42 h, and internucleosomal cleavage was observed by 48 h after oxysterol addition. None of these effects were seen in an oxysterol-resistant CEM subclone, establishing the specificity for apoptosis of the biochemical and morphological events. z-VAD.FMK, a peptide inhibitor of ICE-related proteases delayed but did not prevent the apoptosis of CEM-C7 cells induced by 25OHC. The addition of mevalonate partially protected CEM-C7 cells from apoptosis but did not restore cell growth.

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.

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.

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.

Attenuated fibroblast sheath around the basement membrane zone in the trachea.

In the connective tissue underlying the epithelium of the airways, there are thin attenuated cells that appear to be fibroblasts. These cells are close to the basal lamina and delineate an area equivalent to the basement membrane zone (BMZ). Using the mean data from transversely and longitudinally sectioned attenuated fibroblasts, we estimate that the diameter of the attenuated fibroblast is approximately 28.0 microns and its thickness 0.55 microns. Serial sections indicate there are pores of various sizes in the attenuated fibroblast. Also, the presence of cell segments less than 5 microns in length suggests there are processes from the edge of the cell which would give it a stellate appearance. These large flat cells form a sheath covering 66 to 70% of the BMZ at an average distance of 1.9 microns from the basal lamina. In addition, they make contact with the basal lamina approximately 7,000 times per mm2 of basal lamina. The majority of the contacts with the basal lamina are beneath basal cells. Cells of the attenuated fibroblast sheath are probably associated with lung morphogenesis, wound healing, and development of the BMZ. Their existence as a sheath demonstrates an anatomical unit associated with fibroblast-epithelial cell interactions in airway epithelium.

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)

Differentiation of anchoring junctions in tracheal basal cells in the growing rat.

A function of airway basal cells is to attach ciliated and nonciliated columnar cells to the basal lamina. The significance of the basal cell in attachment is related to the height of the columnar epithelium. In taller epithelia, basal cells are more numerous and differentiated with respect to anchoring junctional adhesion mechanisms (desmosomes, hemidesmosomes, and the cytoskeleton) than in shorter epithelia. In this study, we determined if basal cell anchoring junctional adhesion mechanisms differentiated during growth of the airway. Tracheas from five 3-day-old, five 30-day-old, and five 90-day-old rats were prepared for electron microscopy and morphometrically studied by standard techniques. The circumference of the trachea increased from 2.5 +/- 0.2 to 7.5 +/- 0.4 mm during growth. The height of the columnar cell increased from 13.4 +/- 1.5 to 24.6 +/- 3.9 microns, and the number of basal cells per millimeter increased from 3.2 +/- 0.7 to 9.6 +/- 1.8 during growth. The number of desmosomes per basal cell profile increased significantly from 1.5 +/- 0.1 to 2.1 +/- 0.1, as did keratin filament volume density from 0.046 +/- 0.05 to 0.098 +/- 0.032. The amount of hemidesmosome attachment per basal cell did not increase significantly during growth of the airway. These data demonstrate that as tracheas grow in circumference, the columnar cells increase in height, basal cells increase in number, and anchoring junctional adhesion mechanisms differentiate in the basal cells. These changes are closely related to the height of the epithelium and result in maintaining a constant amount of attachment between the columnar epithelium and the basal lamina as the epithelium increases in height.

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.

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.

Genetic analysis of L-ethionine-mediated induction of alpha-fetoprotein in mice.

Two genetic loci regulate hepatic alpha-fetoprotein (AFP) mRNA levels in adult mice. The raf locus controls basal levels and the Rif locus determines the degree of induction during liver regeneration. We have investigated the function of each locus during L-ethionine-mediated AFP induction using adult female mice with different Rif/raf genotypes. A single intraperitoneal injection of L-ethionine (0.5 mg/g body weight) resulted in significant triglyceride accumulation in hepatic parenchymal cells and increased AFP synthesis 48-96 h following injection. Hepatic AFP mRNA levels in Balb/cJ mice (high basal level/high induction level during regeneration) were 10- to 30-fold higher than Balb/cCRBL or C3H/He mice (low basal level/high induction level) following ethionine injection, indicating that raf-mediated differences persisted throughout the course of acute ethionine poisoning. The magnitude of this induction was similar to that seen during carbon tetrachloride-induced regeneration. In contrast, C57BL/6 mice (low basal level/low induction level during regeneration) contained hepatic AFP mRNA levels similar to Balb/cCRBL and C3H/He mice following ethionine injection. Thus, Rif-dependent differences seen during liver regeneration were not seen during acute ethionine poisoning. This leads us to conclude that (1) hepatic AFP mRNA induction by ethionine may not be mediated by the Rif locus if Rif is a transcriptional inducer, or (2) if Rif is a transcriptional repressor, it is inactivated equally in all strains during acute ethionine poisoning, unlike during liver regeneration. Hepatic albumin mRNA levels were not affected by ethionine treatment in vivo. L-Ethionine elevated AFP mRNA levels in primary mouse hepatocyte cultures; however, ethionine treatment also increased albumin mRNA levels in vitro.

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.

Expression of glutathione S-transferase isoenzymes in human small cell lung cancer cell lines.

A glutathione S-transferase (GST) isoenzyme having common antigenicity to rat placental form (GST-P) and human placental form (GST-pi) has recently been suggested may be a marker of carcinogenesis. In the present study we have investigated the expression of this isoenzyme in three small cell lung cancer cell lines in order to determine whether or not this isoenzyme can be used as a general marker of carcinogenesis. GST activity towards 1-chloro-2,4-dinitrobenzene in two of the cell lines (NES and NOC-361) was moderately higher than that in normal human lung, but this activity was markedly suppressed in one of the cell lines (NCI-H69). Quantitation of the GST isoenzymes in the tumors grown in nude mice by injecting these cell lines also revealed a moderate increase of GST-pi-type isoenzyme in NES and NOC-361 and its suppression in NCI-H69. Immunocytochemical localization studies with these tumors using antibodies raised against GST-pi also indicated a drastic decrease of GST-pi-type isoenzyme in NCI-H69 and this finding was confirmed by Western blot studies. These results suggest that GST-pi, or the isoenzyme(s) having similar immunological nature to GST-pi, cannot be used as the general markers of neoplastic states.