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Translocation of protein tyrosine phosphatase Pez/PTPD2/PTP36 to the nucleus is associated with induction of cell proliferation.

Pez is a non-transmembrane tyrosine phosphatase with homology to the FERM (4.1, ezrin, radixin, moesin) family of proteins. The subcellular localisation of Pez in endothelial cells was found to be regulated by cell density and serum concentration. In confluent monolayers Pez was cytoplasmic, but in cells cultured at low density Pez was nuclear, suggesting that it is a nuclear protein in proliferating cells. This notion is supported by the loss of nuclear Pez when cells are serum-starved to induce quiescence, and the rapid return of Pez to the nucleus upon refeeding with serum to induce proliferation. Vascular endothelial cells normally exist as a quiescent confluent monolayer but become proliferative during angiogenesis or upon vascular injury. Using a 'wound' assay to mimic these events in vitro, Pez was found to be nuclear in the cells that had migrated and were proliferative at the 'wound' edge. TGFbeta, which inhibits cell proliferation but not migration, inhibited the translocation of Pez to the nucleus in the cells at the 'wound' edge, further strengthening the argument that Pez plays a role in the nucleus during cell proliferation. Together, the data presented indicate that Pez is a nuclear tyrosine phosphatase that may play a role in cell proliferation.

Antibodies↗

Labeled Schwann cell transplants versus sural nerve grafts in nerve repair.

This study evaluated the ability of Schwann cell transplants to enhance the recovery of function in injured nerves and compared the results to those produced by sural nerve grafts. Schwann cells were isolated from sciatic nerves, prelabeled with gold fluorescent dye admixed with collagen gel, and placed in resorbable collagen tubes. Twenty-four adult rats underwent severing of the bilateral sciatic nerves, with a 10-mm gap between the nerve stumps. The rats were then divided into two groups. A collagen tube with implanted Schwann cells was implanted in one leg of the Group I rats, and the contralateral leg served as a control and was repaired with a collagen tube filled with collagen gel only. The Group II animals received conduits packed with labeled Schwann cells in one leg to bridge the 10-mm gap; the contralateral leg was repaired with an autogenous sural nerve graft. Recovery of function was assessed physiologically and morphologically. Nerve conduction velocity and nerve action potential amplitude measurements showed that the Schwann cell implants induced return of function comparable to that of the sural nerve grafts. Morphological assessments of myelination suggested a tendency toward greater numbers of myelinated axons in Schwann cell implants than in sural nerve grafts. Anatomical analyses of gold fluorescent dye showed both high viability of prelabeled Schwann cells at 120 days after transplantation and migration as far as 30 mm away from the implant site.

Animals↗

Effects of general anesthesia on myoelectric activity of the intestine in horses.

Myoelectric activity was monitored from the terminal ileum, cecum, and colonic pelvic flexure by use of AgpAgCl bipolar electrodes in 4 adult horses before, during, and after general anesthesia. Horses were anesthetized by way of 3 commonly used regimens, including xylazine (1.1 mg/kg of body weight) and ketamine hydrochloride (2.2 mg/kg); thiopental sodium (7.7 mg/kg), followed by halothane vaporized in oxygen; and thiopental sodium (2.5 g) in guaifenesin (100 mg/ml) solution given to effect, followed by halothane in oxygen. All 3 anesthetic regimens decreased intestinal spike-burst activity in the areas monitored. The slowest return to preanesthetic myoelectric activity was observed after xylazine and ketamine administration. After both of the barbiturate/halothane anesthetic regimens, there was a rebound increase in spike-burst frequency, without alteration in the proportion of propagative myoelectric events. All 3 anesthetic regimens appeared to reset the timing of the small and large intestinal migrating myoelectric complexes. By 9 hours after recovery from anesthesia, the effects of anesthesia, irrespective of regimen, had disappeared. Although anesthesia significantly (P less than 0.05) altered intestinal myoelectric activity, no particular anesthetic regimen had a prolonged effect. Results of our study indicate that the particular chosen regimen of general anesthesia is unimportant in development of motility disturbances in horses after anesthesia.

Acepromazine↗

The effects of ischemia and reperfusion on intestinal motility.

The interdigestive migrating motor complex (MMC) has been demonstrated to be a reliable indicator of intestinal motility and function. The effects of low perfusion on the MMC have never been studied. Fourteen newborn Yorkshire piglets (5 to 18 days old, weighing 2.9 +/- 0.4 kg) underwent celiotomy under general anesthesia with placement of four jejunal electrodes (50 cm apart) as well as a superior mesenteric artery (SMA) Doppler flow probe and a pericardial catheter. Group 1 (n = 5) had operation alone. Group 2 (n = 9) had nonocclusive mesenteric ischemia induced by reversible cardiac tamponade for 5 hours between postoperative days 6 to 12. All subjects had MMC phase III electrical activity, cycling time, and propagation velocity recorded daily. In group 2 MMCs were recorded prior to and during ischemia, and during reperfusion. Group 2 animals had 75% +/- 4% decrease in SMA flow during the tamponade period. During the ischemic period, the MMC cycling time (CT) increased from 67 +/- 10 (mean +/- SEM) to 98 +/- 12 minutes (P < .05) and MMC propagation velocity (PV) decreased to 4.2 +/- 2.2 from a baseline value of 10.5 +/- 1.5 cm/min (P < .05). During reperfusion CT and PV values were not significantly different from baseline. The validity of this model is confirmed by the comparable baseline recordings in groups 1 and 2, and by the return of MMC to baseline values within 4 to 7 hours of reperfusion, as seen in group 2.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Altered motility causes the early gastrointestinal toxicity of irradiation.

PURPOSE: Total abdominal radiation produces symptoms of nausea, vomiting abdominal cramping and diarrhea. Each of these symptoms is associated with disordered intestinal motility. This article reviews studies of large and small intestinal contractile activity following radiation exposure. METHODS AND MATERIALS: Studies of motility utilize strain gauge transducers surgically implanted on the seromuscular layer of the small intestine. All studies were performed in mixed breed dogs to record the occurrence of normal contractions, giant migrating contractions (GMCs) and retrograde giant contractions (RGCs) before, during and after irradiation (22.5 Gy in 9 fractions at 3 fractions/week). Giant migrating contractions and retrograde giant contractions are infrequent in the healthy state. However, in diseased states, GMCs are associated with abdominal cramps and diarrhea, and RGCs precede vomiting. RESULTS: In fasted animals, fractionated abdominal irradiation dramatically increased the frequency of GMCs, with the incidence peaking after the second dose. The increased frequency of GMCS occurred as early as a few hours after the first radiation fraction, and returned to normal within days of cessation of radiation. RGCs were also significantly increased after abdominal irradiation. The frequency of RGCs was greatest on the first and sixth dose of radiation. Clinically, the dogs developed nausea, vomiting and diarrhea as early as the first day of irradiation. In dogs studied in the fed state, decreased amplitude, duration, and frequency of postprandial contractions occurred. These changes may slow intestinal transit during irradiation. Radiation also produced a striking increase in the frequency of colonic GMCs; these changes in colonic motor activity were associated with diarrhea as early as the second irradiation. CONCLUSION: Changes in GI motility during fractionated irradiation precede the appearance of histopathological lesions in the GI tract. Thus, the symptoms of nausea, vomiting, and diarrhea experienced during radiotherapy (particularly those within the first week) are directly related to changes in bowel motility. It is hoped that further understanding of the etiology of these distressing symptoms will help to guide their treatment.

Animals↗

Environmental sodium regulates cutaneous sugar transport in a digenean fluke.

Glucose uptake was examined in adult specimens of Proterometra macrostoma (Trematoda: Digenea) recovered from the stomach (endoparasites) and gills (ectoparasites) of longear sunfish, Lepomis megalotis. The endoparasitic forms transported glucose directly through the external body surface by Na+-independent, facilitated diffusion, but the ectoparasites absorbed glucose by free diffusion alone. To determine how this transport function is regulated, cercariae were incubated in solutions having Na+ concentrations normally found in fish gut (50 mM) and in fresh water (0.5 mM). Glucose transport capacity was retained in 50 mM Na+, but disappeared in worms incubated for 1-3 days in 0.5 mM Na+. Returning worms from the latter solution to one containing 50 mM Na+ fully restored glucose transport within a day. By contrast, incubation up to 5 days in 0.5 mM Na+ had no effect on glucose transport in endoparasitic adults of P. macrostoma and cercariae of P. edneyi. Thus, cutaneous sugar transport function in the migrating larva of P. macrostoma is subject to indirect regulation by environmental Na+.

Animals↗

Pregnancy status determination in mares using a rapid lateral flow test for measuring serum oestrone sulphate.

AIMS: To develop a means of determining pregnancy status in horses based on measuring serum oestrone sulphate (OS) concentrations using a rapid lateral flow immunoassay, and to determine the assay's effectiveness using a visual end-point. METHODS: Serum samples from mares >100 days post-mating (n=701) were assayed using a nitrocellulose membrane-based lateral flow immunoassay device. The device was developed using membrane-bound 1,3,5 (10)-estratrien-3-ol-17-one conjugated to bovine serum albumin as the capture antigen, and an OS-detection monoclonal antibody coupled to colloidal gold as the visible detection reagent. Concentrations of the coating antigen and OS monoclonal antibody were optimised so that the working range would allow pregnancy status to be determined from a visual end-point. The test was run by adding 0.1 ml serum to the sample well of a plastic cassette encasing the test membrane. As the serum migrated along the membrane, a test dot and control line were generated on it within 5-10 min. The intensity of the test dot was inversely proportional to the concentration of OS in the serum sample being tested. Results were compared with those from a validated OS enzyme immunoassay (EIA) and subsequent foaling or return to oestrus of the mares. RESULTS: Serum samples with OS concentrations <10 ng/ml, indicative of non-pregnancy in mares >100 days post-mating, generated a test end-point consisting of a highly visible test dot and control line, whereas serum OS concentrations >50 ng/ml, indicative of pregnancy, generated a control line only. The test correctly identified 384/389 (98.7%) non-pregnant mares tested, and 303/312 (97.1%) pregnant mares tested that were >100 days post-mating. The lateral flow test devices were stable for at least 12 months when stored at 4 degrees C, sealed in aluminium pouches with desiccant. CONCLUSION: This novel, rapid, easy-to-use, lateral flow immunoassay offers a practical alternative to traditional laboratory- based immunoassays for measuring serum OS concentrations in mares for determining their pregnancy status.

Journal Article↗

Behaviour of leatherback sea turtles, Dermochelys coriacea, during the migratory cycle.

Leatherback sea turtles, Dermochelys coriacea, undertake broad oceanic movements. While satellite telemetry has been used to investigate the post-nesting behaviour of female turtles tagged on tropical nesting beaches, long-term behavioural patterns of turtles of different sexes and sizes have not been described. Here we investigate behaviour for 25 subadult and adult male and female turtles satellite-tagged in temperate waters off Nova Scotia, Canada. Although sex and reproductive condition contributed to variation in migratory patterns, the migratory cycle of all turtles included movement between temperate and tropical waters. Marked changes in rates of travel, and diving and surfacing behaviour, accompanied southward movement away from northern foraging areas. As turtles approached higher latitudes the following spring and summer, they assumed behaviours consistent with regular foraging activity and eventually settled in coastal areas off Canada and the northeastern USA. Behavioural patterns corresponding to various phases of the migratory cycle were consistent across multiple animals and were repeated within individuals that completed return movements to northern waters. We consider the potential biological significance of these patterns, including how turtle behaviour relates to predator avoidance, thermoregulation and prey distribution.

Animal Migration↗

The cephalic neural crest provides pericytes and smooth muscle cells to all blood vessels of the face and forebrain.

Most connective tissues in the head develop from neural crest cells (NCCs), an embryonic cell population present only in vertebrates. We show that NCC-derived pericytes and smooth muscle cells are distributed in a sharply circumscribed sector of the vasculature of the avian embryo. As NCCs detach from the neural folds that correspond to the future posterior diencephalon, mesencephalon and rhombencephalon, they migrate between the ectoderm and the neuroepithelium into the anterior/ventral head, encountering mesoderm-derived endothelial precursors. Together, these two cell populations build a vascular tree rooted at the departure of the aorta from the heart and ramified into the capillary plexi that irrigate the forebrain meninges, retinal choroids and all facial structures, before returning to the heart. NCCs ensheath each aortic arch-derived vessel, providing every component except the endothelial cells. Within the meninges, capillaries with pericytes of diencephalic and mesencephalic neural fold origin supply the forebrain, while capillaries with pericytes of mesodermal origin supply the rest of the central nervous system, in a mutually exclusive manner. The two types of head vasculature contact at a few defined points, including the anastomotic vessels of the circle of Willis, immediately ventral to the forebrain/midbrain boundary. Over the course of evolution, the vertebrate subphylum may have exploited the exceptionally broad range of developmental potentialities and the plasticity of NCCs in head remodelling that resulted in the growth of the forebrain.

Animals↗

Intraocular fascioliasis: a case report.

We report the case of a 28-year-old male who had been ill with headache and motor weakness for a month and developed sudden pain and blindness of the right eye. During ophthalmoscopy, a worm was recognized penetrating the iris, occupying the anterior chamber for a brief period, returning back behind the iris, and leaving corneal edema with hyphyema. Enucleation was performed to prevent the worm's escape from the eye. The enucleated eye revealed areas of a focal degeneration of sclera and intraocular hemorrhage. Microscopic findings of an abrupt tissue defect and few inflammatory reactions in the uvea suggested very recent migration of a moving worm. The flatworm detected in the anterior chamber was identified to be a juvenile Fasciola sp. This case is presumed to be the first case of intraocular fascioliasis reported in the literature.

Adult↗

Phenotypic features of smooth muscle cells during the evolution of experimental carotid artery intimal thickening. Biochemical and morphologic studies.

Balloon catheter denudation of rat carotid artery that results in significant medial damage is followed by marked intimal smooth muscle cell (SMC) proliferation associated with limited endothelial regrowth. In this report we demonstrate that: (a) SMC of the carotid media, preceding their intimal proliferation, develop a cytoskeletal profile and morphology consistent with a de-differentiated SMC phenotype; and (b) both medial and intimal SMC subsequently revert to a cytoskeletal profile and morphology reflecting incomplete but significant re-differentiation toward normal SMC phenotype. Specifically, early after balloon injury, SMC of the media and those that have migrated into the intima contain decreased amounts of actin, desmin, and tropomyosin and increased amounts of vimentin; moreover, beta-actin becomes the dominant actin isoform, whereas alpha-actin decreases as compared with that found in normal medial SMC. Late after balloon injury, actin is still less abundant, however, desmin, tropomyosin, and vimentin return toward normal values and both medial and intimal SMC again show a predominance of alpha-actin, although the endothelium does not regenerate over the central surface of intimal thickening in this model. The SMC surface to volume ratio significantly decreases early after balloon injury, whereas it is not significantly different late after balloon injury as compared with that of SMC of the normal carotid media. We demonstrate, furthermore that: (c) adjacent luminal SMC are interconnected by gap junctions and develop focal tight junctions, a feature not reported previously to occur in smooth muscle; these cells however do not form any well defined membrane specialization with the leading edge of endothelium, supporting the view that presence of modified SMC on the luminal surface of chronically denuded vessels is not responsible for the cessation of endothelial regrowth.

Actins↗

[Prurigo and further diagnostically significant skin symptoms in strongyloidosis].

An increasing incidence of strongyloidosis must be expected in European countries as a result of the increasing numbers of immigrants, as well as holiday-makers returning from tropical regions. In addition to gastrointestinal symptoms, dermatological complaints are predominant. Only rarely are cutaneous symptoms the only clinical manifestation of disease. The penetration of filariform larvae may cause "ground itch." In cases of chronic disease, larva currens is the most obvious sign and consists of linear urticarial wheals evoked by larva migration. The most common non-specific symptoms are rashes, pruritus and urticaria. A further symptom of strongyloidosis, intensely itching prurigo, is described in a 20-year-old female Thai. Remission was achieved following tiabendazole therapy.

Adult↗

Migration of donor cells into the thymus is not essential for induction and maintenance of systemic tolerance after liver transplantation in the rat.

In this study, we show that cells encoded by donor-type major histocompatibility complex (MHC) class I antigens effectively migrate into the thymus of the host after liver transplantation. However, the thymus is not essential for the induction and maintenance of tolerance after orthotopic rat liver transplantation (OLT). Liver allografted rats in the DA(RT1a) into PVG (RT1c) combination survived without immunosuppressive agents. The thymus of the host showed a transitory atrophy and returned to normal weight 4 weeks after OLT. Donor antigens were detected in the host thymus after OLT. Adult PVG rats that had been thymectomized 1 week before implantation of DA liver grafts showed liver graft survival without immunosuppression. DA heart grafts were heterotopically transplanted into the PVG thymectomized, liver-grafted rats 1-4 weeks after OLT. No rejection of the cardiac allografts was observed. Systemic tolerance was induced and maintained in the absence of the thymus.

Animals↗

Urokinase-type plasminogen activator receptor reversibly dissociates from complement receptor type 3 (alpha M beta 2' CD11b/CD18) during neutrophil polarization.

Previous studies have shown that the leukocyte integrin CR3 (CD11b/CD18) is physically associated with the urokinase-type plasminogen activator receptor (uPAR;CD87), a glycosyl-phosphatidylinositol (GPI)-linked protein, in resting neutrophil membranes. We now show that uPAR-to-CR3 interactions are reversible, correlating with cell shape. Neutrophils were first labeled with fluorescein conjugates of anti-CR3 F(ab')2 fragments followed by capping using a second-step F(ab')2 directed against murine F(ab')2s. Cells were then probed using rhodamine-conjugated anti-uPAR F(ab')2s. Although uPAR co-caps with CR3 on resting cells, uPAR was found to dissociate or "uncap" coincident with spontaneous cell polarization for migration. CR3 caps transformed into uropods while uPAR accumulated at lamellipodia of polarized cells. Capping was unnecessary for the observed distribution of CR3 and uPAR since the anti-CR3 and anti-uPAR F(ab')2s traffic to the uropod and lamellipodium, respectively, during polarization of uncapped cells. These receptors reassociate when cells return to a spherical morphology. In contrast to uPAR, Fc gamma RIIIB did not dissociate from CR3 caps during cell polarization. Resonance energy transfer (RET) microscopy was used to image the spatial distribution of RET and to follow the kinetics of association and dissociation. Initial levels of RET dramatically fell during cell polarization, but did not change on cells fixed with paraformaldehyde. Receptor reassociation was a biphasic process with initial reassociation about the perimeter of a cap, followed by a plateau and a slower rise in RET within a cap. We suggest that cells regulate receptor-receptor associations depending upon their physiologic activities.

Adult↗

Persistence of foam cells in rabbit xanthoma after normalization of serum cholesterol level.

Xanthoma was produced in diet-induced hypercholesterolemic rabbits by intradermal dextran sulfate injections. The serum cholesterol level returned to the normal range at about 10 weeks after ending the cholesterol diet. Gross observations after cessation of the cholesterol diet revealed a decrease in xanthomatous infiltrations. However, the dense foam cell infiltrations and cholesterol accumulations showed no signs of regression at even 9 months after ending the cholesterol diet. Signs of foam cell migration into the blood stream were not observed. The persistence of the xanthoma may be due to a lack of acceptors, such as high-density lipoproteins, that remove the cholesterol from the foam cells. During our 9-month observation period, some foam cells were degenerated and a few were fused with each other to transform into Touton-type giant cells. Nonfoamy histiocytes were infiltrated around these degenerating foam cells. The histiocytes may have transformed into foam cells by incorporating the lipids of the degenerated foam cells.

Animals↗

Expandable metal stents for the palliation of malignant gastroduodenal obstruction.

PURPOSE: Gastric outlet obstruction is a debilitating complication of upper gastrointestinal malignancy. We present our experience with insertion of self-expanding metal stents (SEMS). METHODS: Twenty-eight patients were referred, stenting being attempted in 23. Two patients had esophageal Wallstents inserted through a gastrostomy; 21 had an endoscopic approach with enteral Wallstents. RESULTS: One stent insertion failed, ten patients (45%) returned to a normal diet, ten patients (45%) managed semi-solid food and two patients (9%) had no significant improvement. No immediate complications were seen. One patient subsequently developed pancreatitis. Reintervention (4 stents, 1 jejunostomy, 1 gastrojejunostomy) was required in six of 22 patients (27%) for inadequate stent expansion (1), second stricture (2), stent migration (1), and tumor ingrowth (2). The mean survival was 95.4 days (SD 78.8 days, range 3-230 days). The mean follow-up time was 98.9 days (SD 86.7 days, range 3-309 days). CONCLUSIONS: SEMS are effective in palliating malignant gastric outlet obstruction. A combined endoscopic/fluoroscopic approach allows the most complete assessment of the stricture and removes the need for gastrostomy insertion. Careful assessment of the gastrointestinal tract distal to the lesion is important.

Adult↗

Differential adaptive response to hyperosmolarity of 3T3 and transformed SV3T3 cells.

Both 3T3 and simian virus 40-transformed 3T3 (SV3T3) cells were used to investigate differences in population kinetics, protein synthesis, monovalent ion levels, and amino acid accumulations between normal and transformed cells exposed to hyperosmolarity at 0.5 Osm. Under similar culture conditions, SV3T3 cells were found to be more sensitive in their proliferative response than normal cells to the hyperosmolar treatment. In the normal 3T3 cells, the increase in transport of amino acids was less sustained and was associated with higher levels of accumulated amino acids. The equilibrium distribution of intracellular monovalent cations and the rate of protein synthesis also returned faster to baseline values in the normal cells than in the transformed cells. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis revealed the induction of a 69-kDa polypeptide in the 3T3 cells but not in the SV3T3 cells after exposure to hyperosmolarity. On electrofocusing and relative mass analysis, this polypeptide closely migrated with the 70-kDa heat shock protein (hsp) family, although it was unrelated immunologically to the inducible 72-kDa hsp.

Amino Acids↗