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Mesothelial progenitor cells and their potential in tissue engineering.

The mesothelium consists of a single layer of flattened mesothelial cells that lines serosal cavities and the majority of internal organs, playing important roles in maintaining normal serosal integrity and function. A mesothelial 'stem' cell has not been identified, but evidence from numerous studies suggests that a progenitor mesothelial cell exists. Although mesothelial cells are of a mesodermal origin, they express characteristics of both epithelial and mesenchymal phenotypes. In addition, following injury, new mesothelium regenerates via centripetal ingrowth of cells from the wound edge and from a free-floating population of cells present in the serosal fluid, the origin of which is currently unknown. Recent findings have shown that mesothelial cells can undergo an epithelial to mesenchymal transition, and transform into myofibroblasts and possibly smooth muscle cells, suggesting plasticity in nature. Further evidence for a mesothelial progenitor comes from tissue engineering applications where mesothelial cells seeded onto tubular constructs have been used to generate vascular replacements and grafts to bridge transected nerve fibres. These findings suggest that mesothelial cell progenitors are able to switch between different cell phenotypes depending on the local environment. However, only by performing detailed investigations involving selective cell isolation, clonal analysis together with cell labelling and tracking studies, will we begin to determine the true existence of a mesothelial stem cell.

Animals↗

Impairment of conjunctival glutathione secretion and ion transport by oxidative stress in an adenovirus type 5 ocular infection model of pigmented rabbits.

Conjunctival epithelial cells of pigmented rabbits secrete reduced glutathione (GSH) into the apical (mucosal) fluid. The aim of the current study was to determine the effect of oxidative stress resulting from viral infection and that of GSH supplementation on redox status, GSH, and ion transport in freshly excised conjunctival tissues and epithelial cell layers in primary culture (RCEC) of adenovirus type 5 (Ad5)-infected rabbits. Lipid peroxidation (LPO) products, nitric oxide (NO), and expression of nitric oxide synthase (NOS2) were quantitated as a function of time after viral inoculation. Unidirectional fluxes of [3H]GSH and changes in short-circuit current (Isc) from mucosal supplementation of Ad5-inoculated conjunctival tissues with GSH and glutathione monoethyl ester (GSH-MEE) were also measured. Ad5 inoculation significantly decreased conjunctival GSH level by 19, 45, 48, and 50% at 8, 24, 48, and 72 h postinfection, respectively. LPO product and NO levels increased significantly (2- and 100-fold, respectively) above that of uninfected controls on Day 3 post-Ad5 inoculation, and co-treatment with GSH-MEE and tocopherol succinate abolished this effect. NO levels showed a progressive increase post-Ad5 inoculation, reaching 0.22 +/- 0.06, 8.12 +/- 0.91, and 2.05 +/- 0.65 microM on Days 1, 3, and 5, respectively, and the highest level was observed on the day of maximal viral replication (Day 3). A very significant induction of the expression of NOS2 on Days 1, 3, and 5 post-Ad5 inoculation was observed. Uninfected control conjunctival tissues displayed a net serosal-to-mucosal GSH flux (Jsm), where the mucosal-to-serosal flux (Jms) was approximately 14 pmol h(-1) cm(-2) and the Jsm was approximately 22 pmol h(-1) cm(-2). In Ad5-inoculated rabbits similar GSH flux was observed in both the sm and ms directions, and the net GSH flux was negligible. Isc and potential difference (PD) across conjunctival tissues of Ad5-inoculated rabbits decreased by > or = 50% compared with control, while the transepithelial electrical resistance (TEER) remained unchanged. Mucosal, but not serosal, superfusion of GSH or GSH-MEE in Ad5-inoculated conjunctival tissues increased the Isc by up to 40% in approximately 100 min. Our results show that net secretion of GSH across rabbit conjunctiva is totally blocked after Ad5 inoculation and active ion transport rate decreased by approximately 50%. Decreased net GSH secretion into mucosal fluid after Ad5 infection may have resulted from a decreased intracellular GSH pool due to oxyradical-induced changes in redox status and lower active ion transport. Mucosal treatment of Ad5-infected conjunctival tissues with pharmacological levels of GSH appears to transstimulate mucosal GSH secretion and restore active ion transport activity, suggesting a potentially useful therapeutic regimen for ocular infections.

Adenoviridae↗

Development of embryonic membranes in the silverfish Lepisma saccharina linnaeus (insecta: Zygentoma, Lepismatidae).

Development and fate of embryonic membranes in the silverfish Lepisma saccharina was examined throughout embryogenesis. The amnioserosal folds first arise as serosal folds that are completed by the later addition of the amnion from the embryo's margins as in archaeognaths. The close link between production of the amnion and formation of the folds should not be assigned to Dicondylia but to Pterygota as an autapomorphy. During fold formation, folding of embryonic membranes beneath the embryo is less extensive and the ventral cupping of the embryo plays a larger role comparable to that occurring in archaeognath embryos. In L. saccharina, the embryonic membrane pore (the amniopore) varies in its manner of closure, either by complete fusion of serosal folds or by formation of a serosal cuticular plug between them as in archaeognaths. Although, in many aspects of its embryogenesis, L. saccharina retains the primitiveness of archaeognaths, its amnioserosal folds persist and are well integrated into its embryogenesis as the amnioserosal fold-amniotic cavity system is established and as occurs in many pterygote embryos; this may be thus regarded as an autapomorphy of Dicondylia.

Animals↗

Absorption of quercetin-3-glucoside and quercetin-4'-glucoside in the rat small intestine: the role of lactase phlorizin hydrolase and the sodium-dependent glucose transporter.

Two hypotheses on absorption mechanisms of flavonoid glucosides across the small intestine have been proposed: active uptake of the quercetin glucoside by the sodium-dependent glucose transporter (SGLT1) with subsequent deglycosylation within the enterocyte by cytosolic beta-glucosidase, or luminal hydrolysis of the glucoside by lactase phlorizin hydrolase (LPH) and absorption by passive diffusion of the released aglycone. To test the above hypotheses we employed phlorizin (as an inhibitor of SGLT1) and N-(n-butyl)-deoxygalactonojirimycin (as an inhibitor of the lactase domain of LPH) in a rat everted-jejunal sac model. Quercetin-4'-glucoside mucosal hydrolysis was 10 times greater than quercetin-3-glucoside hydrolysis in the absence of inhibitors (449 and 47 nmol g(-1) tissue, respectively), despite the similar amounts (13+/-4 and 9+/-1 nmol g(-1), respectively) being transferred to the serosal compartment during the 15 min incubation. Apical hydrolysis of both quercetin glucosides was significantly reduced in the presence of NB-DGJ (80%), and transfer of quercetin (measured as quercetin metabolites) to the serosal solution was also significantly reduced (40-50%). In the presence of phlorizin, transfer of metabolites to the serosal solution was only reduced in the case of quercetin-4'-glucoside. Evidently the mechanism of absorption of quercetin-4'-glucoside involves both an interaction with SGLT1 and luminal hydrolysis by LPH, whereas quercetin-3-glucoside appears to be absorbed only following hydrolysis by LPH.

Analysis of Variance↗

Gossypol induces chloride secretion in rat proximal colon.

The effect of gossypol on electrolyte transport was investigated in rat colon mounted in Ussing chambers. The addition of gossypol to the mucosal or serosal side led to an increase in mucus secretion, which we did not quantify. Mucosally or serosally added gossypol also induced a rise in short circuit current (I(sc)) and tissue conductance (G(t)). Part of the mucosally added gossypol seemed to be bound to the mucus because the effects on I(sc) and G(t) were smaller when gossypol was added to the mucosal side. Serosally added gossypol had an effect on I(sc) at a concentration of 10 micromol l(-1). Mucus secretion was reduced in low Ca(2+) buffer. The increase in I(sc) was diminished by blockers of Cl- channels, K+ channels, of the Na+/K+ ATPase and of the Na+/K+/2 Cl- cotransporter. Measurements of unidirectional ion fluxes showed that gossypol added to the mucosal side had no effect on net Na+ transport, but increased Cl- secretion. The effect of mucosally added gossypol was significantly reduced by the use of low Cl- buffers and abolished when the buffer was additionally depleted of HCO(3)(-). Calmodulin antagonists inhibited the effect on secretion. These findings indicate that gossypol induces chloride secretion via a calmodulin-dependent mechanism. High concentrations of gossypol induced a strong increase in G(t) that could be blocked by W7, a blocker of calmodulin-dependent myosin light chain kinase. This indicates that the rise in G(t) is not due to an unspecific toxic effect, but instead, to specific opening of tight junctions.

Animals↗

Expression of transforming growth factor beta isoform mRNA in injured peritoneum that healed with adhesions and without adhesions and in uninjured peritoneum.

OBJECTIVE: To compare the mRNA levels of transforming growth factor beta (TGF-beta) isoforms 1, 2, and 3 among surgically injured peritoneum that healed with adhesions, surgically injured peritoneum that healed without adhesions, and uninjured peritoneum. DESIGN: Prospective experimental study. SETTING: University vivarium. ANIMAL(S): Fourteen sexually mature female Sprague-Dawley rats, 226-250 grams. INTERVENTION(S): Standardized cecal abrasion was performed on 120 Sprague-Dawley rats. The areas of abrasion and any resultant adhesions were harvested at necropsy 7 days later. Total RNA was then extracted from the serosal adhesions of four rats, from the serosa of five rats that healed without adhesion formation, and from analogous areas of cecum from five rats that did not undergo abrasion and served as controls. MAIN OUTCOME MEASURE(S): Quantification of the expression of the TGF-beta1, TGF-beta2, and TGF-beta3 mRNA transcripts in peritoneal adhesions, normally healed peritoneum, and fresh, uninjured peritoneal tissue through the use of multiplex reverse transcriptase-polymerase chain reaction analysis. RESULT(S): Peritoneal adhesion TGF-beta1 and TGF-beta3 mRNA expression was 3.7-fold and 8.6-fold higher, respectively, than in abraded tissues that healed normally; and 5.6-fold and 4.6-fold higher, respectively, than in nonabraded tissues. While TGF-beta2 mRNA levels were also higher in serosal adhesions compared with normally healed and uninjured peritoneum, this rise was not statically significant. CONCLUSION(S): Peritoneal adhesions resulting from surgical abrasion of a serosal surface have statistically significant increased levels of TGF-beta1 and -beta3 mRNA transcripts compared with both uninjured and normally healed peritoneum.

Animals↗

Evaluation of laser Doppler flowmetry in the assessment of intestinal blood flow in cat.

Laser Doppler flowmetry with a differential detector system has been used in the assessment of blood flow in the feline small intestine. Simultaneous mucosal and serosal laser Doppler flowmeter recordings were compared with total blood flow of a bowel segment measured by an optical drop-recorder unit in 6 cats. Blood flow through the muscularis layer was estimated using the 85Kr washout technique. A correlation coefficient of r = 0.96 (mucosal recordings = 90, serosal recordings = 80, p less than 0.001) was obtained between laser Doppler flowmeter output signals and total blood flow at different levels of vascular tone, regardless of whether the flowmeter recordings were made from the mucosal or the serosal side of the bowel wall. At intense vasodilation, the flowmeters showed a tendency to underestimate blood flow. The flowmeter signals were at variance with the muscularis blood flow but were clearly correlated to the calculated mucosal-submucosal blood flow. The uneven blood flow distribution in the intestinal wall did not affect the ability of the flowmeters to reflect total blood flow from either side of the bowel wall. A calibration curve could be constructed for approximate interpretation of the laser Doppler signals in absolute flow units. However, further experiments in humans and further development of the technique must be performed to elucidate clinical applications of the method.

Animals↗

Hemostatic effect of local intramural injection of dehydrated ethanol in the canine gastrointestinal tract.

The relative value of a subserosal injection of 98% ethanol (0.2 ml x 4) in controlling acute and chronic bleeding from serosal vessels was assessed in 17 dogs. Blood flow was measured from gastric serosal vessels (average diameter, 1.6 mm) severed immediately after, 24 hours after, and 48 hours after ethanol injection. Blood flow from severed colonic serosal vessels (averaging diameter, 1.0 mm) was measured before and immediately after ethanol injection. The safety of ethanol injection was tested by endoscopically guided submucosal injections which were sequentially observed by endoscopy at 1 hour, 24 hours, and weekly for 4 weeks after injection. Ethanol injection had no effect on bleeding from larger gastric vessels unless the injection was made 24 or 48 hours prior to vessel severence. Ethanol injection was effective in reducing bleeding from the smaller colonic vessels when done immediately prior to vessel severence. Gastric submucosal injections led to ulcers which extended into the muscle layer at 1 week and were completely healed by 3 weeks; none perforated or bled. These data support the potential efficacy of therapeutic ethanol injection for the control of small vessel (1.0 mm in diameter) bleeding and the potential prophylactic value against rebleeding from larger vessels. Further studies are needed to determine whether these findings are organ related as opposed to being diameter specific.

Animals↗

Seromuscular enterocystoplasty in rats.

Enterocystoplasty is commonly used in clinical practice. Many of its undesirable effects, that is infections, stones, mucus production, absorption of urinary components into the blood stream and risk of cancer, result from the intestinal mucosa lining the urinary tract. We report on the feasibility of creating an enterocytoplasty with a seromuscular colonic segment that acquires a transitional epithelial lining. Augmentation enterocystoplasty was performed in 51 male, 500 gm., Sprague-Dawley rats with a 1.82 cm.2 patch of left colon from which the mucosa had been stripped. The serosal surface was used as lining for the enterocystoplasty. The intestinal patch and the bladder capacity at known pressure were measured at operation and at sacrifice. The histology of the enterocystoplasty was studied in detail following sacrifice. Of the animals 40 survived without significant complications and were sacrificed at a mean postoperative time of 30 days (range 5 to 80 days). In the remaining 11 rats a bladder stone developed but it did not seem to affect the outcome of the experiment. The size of the patch could be measured in 22 rats: it was 1.82 cm.2 (standard deviation +/- 0.86) at operation and 2.30 cm.2 (standard deviation +/- 1.1) at sacrifice. In none of the rats did the patch decrease in size. The bladder capacity at a known pressure (mean 17 cm. water) could be measured in 26 animals: it was 2.35 ml. (standard deviation +/- 0.65) at operation and 5.18 ml. (standard deviation +/- 1.19) at sacrifice. Histological analysis was done in 40 rats. In all cases the serosal surface was lined with transitional epithelium, there was no fibrosis or inflammation and the structure of the muscular layer of the bowel remained intact. The earliest growth of uroepithelium in the serosal surface of the bowel was noted at 5 days. This model suggests that the seromuscular enterocystoplasty can be constructed successfully in the rat model. The seromuscular intestinal patch does not shrink. The bladder capacity increases and histology shows a uroepithelial lining of the augmentation.

Animals↗

Protein kinase C regulates prairie dog gallbladder ion transport.

BACKGROUND: Gallstone formation is characterized by increased biliary calcium (Ca2+) level and altered gallbladder absorption. Recent studies suggest that luminal Ca2+ regulates gallbladder ion transport via intracellular calcium ([Ca2+]ic). Ca2+-calmodulin and protein kinase C (PKC) are two major systems through which [Ca2+]ic carries out second-messenger functions in many cell types. We have previously shown that Ca2+-calmodulin regulates basal gallbladder ion transport in prairie dog. The present study tests the hypothesis that PKC is also essential in regulation of gallbladder ion transport in this model. METHODS: The role of PKC in regulation of gallbladder ion transport was determined by studying the effects of phorbol esters, synthetic analogues of diacylglycerol, which directly activates PKC. Gallbladders were mounted in Ussing chambers, and standard electrophysiologic parameters were recorded after exposing tissues to either 10(-5) mol/L of 4-alpha-phorbol 12,13-didecanoate (PDD), 4-beta-phorbol 12-myristate 13-acetate, 4-beta-phorbol 12,13-dibutyrate (PDB), or 10(-4) mol/L serotonin. Unidirectional Na+, Cl-, and H2O fluxes were measured before and after treatment with only inactive PDD and most active PDB. RESULTS: Mucosal and serosal exposure of tissues to either 4-beta-phorbol 12-myristate 13-acetate or PDB resulted in a decrease in short-circuit current and transepithelial potential difference without any change in tissue resistance. Serotonin induced similar changes in gallbladder electrical properties. PDB caused an inhibition of mucosal to serosal fluxes of Na+, Cl-, and H2O, with a decrease in net Na+ absorption, an increase in net Cl- secretion, and a conversion of net H2O absorption to net H2O secretion. Serosal-to-mucosal fluxes of Na+, Cl-, and H2O did not change. Inactive PDD had no effect on either electrophysiologic parameters or ion and water fluxes. Pretreatment of tissues with PKC antagonist 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine blocked the phorbol ester-induced inhibition of ion transport. CONCLUSION: PKC regulates gallbladder ion transport in the prairie dog by inhibiting Na+ absorption and stimulating Cl- secretion.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Ovicide-induced serosa degeneration and its impact on embryonic development in Manduca sexta (Insecta: Lepidoptera).

Eggs of Manduca sexta treated with the ovicide Ov. 165049 turn orange, and the embryos later die. The orange pigmentation is at first confined to the serosa, and is accompanied by pathological changes of serosal cells. Lipid vesicles aggregate and spindle-shaped electron-lucent vesicles-normally forming a single layer below the apical cell surface-greatly accumulate. The mitochondria swell considerably, and their matrices become electron-lucent. Subsequently, the serosal cells develop additional features of necrosis. They form many autophagic vacuoles which contain mostly degradating mitochondria, but also segregated rough endoplasmic reticulum (rER) and glycogen granules. The whole cytoplasm vesiculates, and the cells shrink considerably. The nuclei become less irregular in shape, the chromatin disperses rather evenly whereas the nucleoli persist. Neither chromatin condensation nor the production of apoptotic bodies was observed-further evidence, that the serosal cells die by necrosis rather than apoptosis. At some stage of development the damaged serosa ruptures, retracts from the embryo and forms a sphere beneath it. It is only after the rupture of the serosa, that the embryo also turns orange and disintegrates rapidly. This shows impressively the protective function which the serosa plays for the embryo. Our physiological tests indicate, that the orange pigmentation of the serosa induced by the ovicide results from a disturbance of the tryptophan/ommochrome pathway serving the excretion of potentially toxic metabolites of tryptophan-rich proteins. The results demonstrate first that the serosa represents an important target for ovicide pesticides and second that it plays a vital role as an excretory organ during embryogenesis.

Animals↗

Intestinal transport and metabolism of glucose-conjugated kyotorphin and cyclic kyotorphin: metabolic degradation is crucial to intestinal absorption of peptide drugs.

Intestinal transport and metabolism of modified kyotorphin (KTP) were studied in rats. Modified KTPs studied were C-terminally modified KTP with p-aminophenyl-beta-D-glucoside (KTP-pAPbeta glc), N-terminally modified KTP-pAPbeta glc with t-butyloxycarbonyl group (Boc-KTP-pAPbeta glc) and the N- and C-terminally modified KTP by cyclization (cyclic KTP). KTP-pAPbeta glc was metabolized at a similar rate to that of KTP, and did not appear on the serosal side. Although Boc-KTP-pAPbeta glc was also metabolized, it was more stable than KTP and appeared on the serosal side. Cyclic KTP was also quite stable and appeared on the serosal side. The modified KTPs were evaluated kinetically for absorption consisting of membrane transport and metabolism. Absorption clearance (CL(abs)) of cyclic KTP, Boc-KTP-pAPbeta glc and Boc-KTP was higher than that of KTP (0.247 microl/min/cm) (Mizuma et al., Biochim. Biophys. Acta 1335 (1997) 111-119), which is the theoretical maximum by complete inhibition of peptidase activity, indicating that derivatization of KTP increases the membrane permeability. Furthermore, the data clearly showed that the greater the metabolic clearance (CL(met)) of KTP and the KTP derivatives, the lower the absorption clearance (CL(abs)). These results and further simulation study led to the conclusion that metabolic degradation in the intestinal tissues is more critical than membrane permeability (transport) for oral delivery of peptide drugs. Based on the stability of cyclic KTP in serum, this appears to be a good candidate analgesic peptide drug.

Acetaminophen↗

Intestinal transport and metabolism of analgesic dipeptide, kyotorphin: rate-limiting factor in intestinal absorption of peptide as drug.

Intestinal transport and metabolism of kyotorphin (KTP) were studied in rat everted small intestine. KTP on the mucosal side was metabolized completely within 60 min, and any amounts of KTP were not detected on the serosal side. On the other hand, [D-Arg2]-KTP (D-KTP) was stable on the mucosal side to appear on the serosal side. However, N-t-butoxycarbonyl-KTP (Boc-KTP), which was metabolized on the mucosal side faster than KTP, appeared on the serosal side. In intestinal homogenate, KTP was metabolized, and the metabolic clearance (CL(met)) was decreased by peptidase inhibitors, bestatin, o-phenanthrolin and tryptophan hydroxamate. In the presence of these peptidase inhibitors, the absorption clearance (CL(abs)) of KTP was increased. The less the CL(met) of KTP was, the more the CL(abs) of KTP was. Meanwhile, Boc-KTP in intestinal homogenate was stable even in the absence of peptidase inhibitors. The CL(abs) of Boc-KTP was constant irrespective of the stability on the mucosal side. Kinetic analysis by the metabolic inhibition model indicated that the stabilization of KTP in the intestinal tissue could increase the CL(abs) up to 0.247 microl/min per cm, which was as much as the CL(abs) of stable D-KTP. These results led to the conclusion that rate-limiting process in intestinal absorption of KTP is metabolic degradation in intestinal tissue during the absorption.

Analgesics↗

Projections of pelvic autonomic neurons within the lower bowel of the male rat: an anterograde labelling study.

The tissues of the large intestine which receive an innervation by neurons of the major pelvic ganglia were identified following in vivo and in vitro anterograde labelling with the lipophilic tracer 1,1'didodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate in the male rat. The primary target in the gut of major pelvic ganglion neurons is the myenteric plexus of the distal colon and the rectum. The serosal ganglia, on the surface of the most distal region of the rectum and the circular muscle of the distal colon and rectum were less densely innervated. The pelvic ganglia do not innervate the longitudinal muscle, submucosal blood vessels, submucosal plexus, or mucosa. The pelvic supply reaches the bowel via two groups of rectal nerves and branches of the penile nerves. All of these connections also carry the axons of viscerofugal neurons from the bowel, some of which have terminal axons in the major pelvic ganglia. Finally, the different nerves supplied different targets. In particular, while the rectal nerves carried pelvic axons supplying the myenteric plexus, circular muscle, and serosal ganglia, the penile nerves only innervated the serosal ganglia. In addition, the two groups of rectal nerves innervated slightly different regions of the bowel and provided different projection patterns. However, successful in vivo labelling was achieved in only 6/12 animals and while all in vitro experiments resulted in successful labelling, it was clear that only a proportion of pelvic projections in any given nerve were labelled. These studies have shown that the major pelvic ganglia are primarily involved in the control of motility, but not of vascular and secretomotor functions. Thus pelvic neurons do not innervate the same range of target tissues within the bowel as the prevertebral ganglia. This study has also shown that the different pathways to the gut from the major pelvic ganglia innervate different tissues, suggesting that the autonomic innervation of the gut is not homogeneous along its length.

Animals↗

Arginine vasopressin transport and metabolism in the pigmented rabbit conjunctiva.

The purpose of this study was to evaluate the transepithelial transport and metabolism of arginine vasopressin (AVP) in the pigmented rabbit conjunctiva, both in the absence and presence of protease inhibitors. The apparent permeability coefficient, P(app), for 3H-AVP was determined in the modified Ussing chamber, and AVP metabolites were monitored by reversed phase HPLC using a C18 column. At 50 nM donor 3H-AVP, the P(app) in the mucosal-to-serosal (ms) direction was about five times higher than that in the opposite direction. Excess (0.1 mM) AVP decreased the P(app) for labelled AVP in the mucosal-to-serosal (ms) direction by about 50%. However, intact AVP transport showed neither concentration nor direction dependence. HPLC analysis revealed two subspecies of 3H-AVP in the receiver fluid and virtually no degradation products in the donor fluid following 3 h flux experiments. 3H-AVP transported in the ms direction underwent extensive hydrolysis (73%), which was decreased by 33% with mucosal application of 2 mM camostat mesylate (an aminopeptidase inhibitor) or by 27% with 0.5 mM leupeptin (a serine protease inhibitor). By contrast, 3H-AVP transported in the serosal-to-mucosal (sm) direction resulted in only 37% hydrolysis, and mucosal application of either inhibitor did not significantly affect the P(app) for intact AVP. These data suggest that intact AVP transport in the conjunctiva may be mediated mostly by passive diffusion and enzymatic degradation of AVP may be mediated by proteolytic enzymes present on the mucosal side of the conjunctiva.

Animals↗

Dosimetric considerations in the treatment of inoperable endometrial carcinoma by a high dose rate afterloading packing technique.

A series of 23 patients with early adenocarcinoma of the endometrium who underwent a total of 37 modified Heyman packings treated on a high dose rate Microselectron has been reviewed. Using computed tomography (CT), the uterine wall thickness was measured retrospectively and doses calculated at a number of points on the uterine serosa and related normal tissues. The mean and maximum fundal serosal doses were found to be highest posteriorly and the sigmoid colon was adjacent to the posterior surface of the uterus in all instances. By superimposing the isodose distribution on CT sections of the uterus, it is now possible to prescribe to a serosal dose, or, in patients too heavy for the CT scanner, a dose can be prescribed to a point S, which is a reasonable approximation to the serosal position. Since the initial study, a further ten patients have been treated by the same method and, where relevant, data from all 33 patients have been used.

Adenocarcinoma↗

pH and energy dependent transport of ketoprofen across rat jejunum in vitro.

The aim of this study was to elucidate transport mechanisms of ketoprofen (monocarboxylic acid with pK(a) 4.6) across rat jejunum in vitro using side-by-side diffusion cells. When the tissue was incubated on the mucosal and serosal sides with buffer of pH 7.51 (pH of the mucosal surface was 7.08), ketoprofen permeated faster in the mucosal-to-serosal than in the opposite direction. No asymmetry in transport was observed when 2 mM mucus disrupting agent 1,4-dithio-DL-threitol (pH of the mucosal surface increased to 7.21) was added to the mucosal side. Mucosal-to-serosal permeability of ketoprofen increased three times when the pH of the incubation medium was changed from 8.06 (pH of the mucosal surface was 7.34) to 6.07 (pH of the mucosal surface was 5.95), while no pH dependence was found under ATP-depletion caused by sodium azide. In the ketoprofen concentration range from 0.125 to 5 mM no saturation of transport was observed. Moreover, ketoprofen transport was not changed in the presence of 2 mM benzoate, 10 and 20 mM acetate, 20 mM L-lactate (substrates for monocarboxylate transporter 1, MCT1) and 1 mM alpha-cyano-4-hydroxy-cinnamic acid (an inhibitor of MCT1). These results indicate that ketoprofen is transported across rat jejunum in vitro by pH and energy dependent transport mechanisms, and most probably not by MCT1.

Adenosine Triphosphate↗

A novel method for chronic measurement of respiratory function in the conscious monkey.

INTRODUCTION: Chronic measurement of respiratory function in the conscious monkey has been limited by the ability to monitor pleural pressure. Previous attempts to measure pleural pressure chronically in conscious animals have involved surgical implantation of pressure-sensitive catheters directly into the pleural cavity. The success of these techniques has been limited by lung damage and/or tissue growth and encapsulation of the pressure-sensitive catheter with damping or loss of the signal. These problems have been eliminated by developing a novel surgical procedure for placement of a pressure-sensitive catheter beneath the pleural surface. METHODS: A pressure-sensitive catheter (attached to a radiotelemetry transmitter) was surgically implanted beneath the serosal layer of the esophagus within the thoracic cavity. This was accomplished by performing a midline laparotomy, making a small incision in the serosal layer of the esophagus caudal to the diaphragm, and advancing the catheter cranially along the esophagus and beneath the serosal layer into the thoracic cavity. The catheter was secured to the esophageal wall and the telemetry transmitter was sutured to the inner abdominal wall. Respiratory airflow measurements were obtained using a restraint chair equipped with a clear plastic helmet (plethysmograph chamber) that seals around the neck and encloses the head. RESULTS: The decreases in pleural pressure during inspiration ranged from -5 to -15 mmHg and remained constant for at least 36 weeks following surgery. Intravenous administration of the respiratory depressant morphine, exposure to the respiratory stimulant 8% CO2, and intravenous administration of the bronchoconstrictive agent methacholine verified that this technique was able to detect and quantify ventilatory changes and an increase in airway resistance. DISCUSSION: This novel method for chronic measurement of pleural pressure will facilitate evaluation of the effects of drugs, environmental agents, or disease on respiratory function by allowing repeated measurements of both ventilatory (breathing) patterns and the mechanical properties of the lung in a conscious nonhuman primate.

Airway Resistance↗