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J Yu

Publications and source records attributed to J Yu.

At least 811 records · Page 45Linked to original sources

Large scale purification and immunolocalization of bovine uroplakins I, II, and III. Molecular markers of urothelial differentiation.

The differentiation of mammalian urothelium culminates in the formation of asymmetrical unit membrane (AUM). Using gradient centrifugation and detergent wash, we purified milligram quantities of AUMs which, interestingly, contained three major proteins (15, 27, and 47 kDa) that appeared to be identical to the three immunoaffinity purified, putatively AUM-associated proteins that we described earlier (Yu, J., Manabe, M., Wu, X.-R., Xu, C., Surya, B., and Sun, T.-T. (1990) J. Cell Biol., 111, 1207-1216). Peptide mapping and immunoblotting established that these three proteins were distinct molecules. Using monospecific antibodies to these three proteins, we showed that they were all restricted to the superficial urothelial cells and were AUM-associated. The 27- and 15-kDa proteins were detected exclusively on the luminal side of mature, apical AUMs. In contrast, epitopes of the 47-kDa protein were detected on both sides of apical AUMs suggesting a transmembranous configuration. These results (i) provide the strongest evidence thus far that AUM contains three major proteins (the 27-kDa uroplakin I, 15-kDa uroplakin II, and 47-kDa uroplakin III) which form an extremely insoluble complex, (ii) suggest that uroplakin II, like uroplakin I (Yu, J., Manabe, M., Wu, X.-R., Xu, C., Surya, B., and Sun, T.-T. (1990) J. Cell. Biol. 111, 1207-1216), translocates from one side of the membrane to another during AUM maturation, (iii) indicate that uroplakin III may play a different structural role than uroplakins I and II in AUM formation, and (iv) establish the three uroplakins as markers for an advanced stage of urothelial differentiation.

Animals↗

Neural systems contributing to acetylcholinesterase histochemical staining in primary visual cortex of the adult rat.

Histochemical studies demonstrate that cortical area 17 (primary visual cortex) of the adult rat displays a characteristic laminar pattern of acetylcholinesterase (AChE) activity. While AChE-positive axons are found throughout the cortical layers, most intense staining occurs in a band that corresponds to layer V and the deep portion of layer IV. The present studies were directed toward determining the neural systems containing this AChE activity. Unilateral electrolytic or excitatory amino acid induced lesions of the basal forebrain result in reductions of AChE staining in ipsilateral visual cortex, particularly in layers IV and V. Electrolytic or scalpel lesions, placed in white matter underlying dorsal and lateral neocortex to interrupt basal forebrain projections to visual cortex, also reduce AChE staining in visual cortex. Lesions in the cingulate bundle and supracallosal stria reduced AChE staining retrosplenial cortex but did not affect staining visual cortex. Placement of electrolytic lesions in the hypothalamus produced no detectable change in the pattern of AChE in visual cortex. Electrolytic lesions in the midbrain tegmentum, placed to interrupt ascending axons from brainstem monoamine neurons, produced no detectable change in the pattern of AChE in visual cortex. Placement of lesions in the dorsal thalamus that include all of the dorsal lateral geniculate nucleus did not alter AChE staining in visual cortex. The results indicate that AChE activity in adult visual cortex is found primarily within afferent axons from the basal forebrain system. These data demonstrate further that the AChE staining characteristic of adult visual cortex is associated with neural systems that are distinctly different from those associated with AChE staining in visual cortex of the infant rat.

Acetylcholinesterase↗

Transfusion-induced immunosuppression results in diminished host survival in a murine neuroblastoma model.

Perioperative blood transfusion has been associated with decreased survival in cancer patients. The immunologic consequences of H-2 incompatible blood transfusion as related to neoplasia are unclear. This report examined the effect of multiple allogeneic blood transfusions, compared to syngeneic transfusions and saline infusion, on cellular immunity, tumor growth, and host survival in a murine C1300 neuroblastoma model. A/J mice were randomized to receive two weekly transfusions of washed whole blood cells from C57 Bl/6 or A/J donors or saline. Animals transfused with allogeneic blood, compared to syngeneic transfusions or saline infusions, had a significantly diminished lymphocyte response to mitogen (P less than 0.001), reduced donor-specific (P less than 0.001) and third party alloantigen (P less than 0.01) MLR, and reduced cytotoxicity against a natural killer (NK) cell-sensitive target (P less than 0.001). These in vitro deficits in cellular immunity correlated with a significantly greater Day 21 tumor weight to total body weight ratio in the allogeneic group (0.33) compared with the syngeneic (0.25) and saline (0.28) groups P less than 0.05). Median host survival was reduced in the allogeneic group (24 days) compared with the syngeneic (30 days) and saline (31 days) groups. There were no significant differences in cellular immunity, tumor growth, or survival between syngeneic and saline control groups. Allogeneic blood transfusion had an adverse affect on NK and T-lymphocyte function which was associated with enhanced tumor growth and reduced survival in tumor-bearing mice.

Animals↗

Indirect effects of histamine on pulmonary rapidly adapting receptors in cats.

We did experiments to determine the relative importance of lung mechanical changes during histamine induced activation of pulmonary rapidly adapting receptors (RARs). In anesthetized, open-chest, artificially ventilated cats, we recorded RAR activity and injected histamine (25-50 micrograms/kg) into the right atrium. Histamine initially increased RAR activity from 1.1 +/- 0.2 to 3.6 +/- 0.6 imp/sec (n = 30) at 15.6 +/- 0.8 sec when dynamic lung compliance (CDYN) was decreased by 29.1 +/- 1.5%. The firing pattern of RARs changed from a relatively irregular pattern to a pronounced respiratory modulation. RAR activity reached its peak (5.6 +/- 0.8 imp/sec) at 36.3 +/- 3.3 sec. The firing pattern further changed to a cardiac modulation, and the activity closely correlated with cardiac output. Comparing the initial response of RARs to histamine with the response to mechanically decreasing CDYN, we found that the activities were similar when CDYN was decreased by the same amount. Our experiments suggest that in cats the initial increase of RAR activity in response to histamine is related to lung mechanical changes, but the later increase is related to cardiovascular functions.

Action Potentials↗

Patterns of transiently expressed acetylcholinesterase activity in cerebral cortex and dorsal thalamus of developing rats with cytotoxin-induced microencephaly.

Previous studies have demonstrated that acetylcholinesterase (AChE) activity is expressed transiently by thalamocortical neurons of primary sensory systems in developing rat pups. In the present study, prenatal treatment with methylazoxymethanol acetate (MAM) on embryonic day 15, 16, or 17 resulted in rat pups with cerebral cortices markedly reduced in thickness and areal extent. Histochemical studies demonstrated that AChE staining occurs in fiber-like plexuses in primary visual, auditory, and somatosensory regions of developing cerebral cortex of MAM-treated animals, just as in normal developing rats, but that the transient patterns of AChE are found more superficially than normal and they occur in an abnormal patchy distribution. Neuronal somata in thalamic lateral geniculate, medial geniculate and ventral basal nuclei of MAM-treated animals show transient AChE staining indistinguishable from that seen in normal animals. These data indicate: (1) AChE is expressed transiently by thalamocortical neurons in MAM-treated animals, (2) intensity of the transiently expressed AChE is not affected by MAM-induced loss of cortical neurons, and (3) the abnormal AChE patterns in cortex likely reflect the abnormal distributions of thalamocortical terminal fields that are characteristic of MAM-treated animals.

Acetylcholinesterase↗

A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells.

The ability of enteropathogenic Escherichia coli (EPEC) to form attaching and effacing intestinal lesions is a major characteristic of EPEC pathogenesis. Using TnphoA mutagenesis we have identified a chromosomal gene (eae, for E. coli attaching and effacing) that is necessary for this activity. A DNA probe derived from this gene hybridizes to 100% of E. coli of EPEC serogroups that demonstrate attaching and effacing activity on tissue culture cells as well as other pathogenic E. coli that produce attaching and effacing intestinal lesions, such as RDEC-1 (an EPEC of weanling rabbits) and enterohemorrhagic E. coli. The predicted amino acid sequence derived from the nucleotide sequence of eae shows significant homology to that of the invasin of Yersinia pseudotuberculosis.

Animals↗

Uroplakin I: a 27-kD protein associated with the asymmetric unit membrane of mammalian urothelium.

The luminal surface of mammalian urothelium is covered with numerous plaques (also known as the asymmetric unit membrane or AUM) composed of semi-crystalline, hexagonal arrays of 12-nm protein particles. Despite the presumed importance of these plaques in stabilizing the urothelial surface during bladder distention, relatively little is known about their protein composition. Using a mouse mAb, AE31, we have identified a 27-kD protein that is urothelium-specific and is differentially expressed in superficial umbrella cells. This protein (pI approximately 5.8) partitions into the detergent phase during Triton X-114 phase separation. Pulse-chase experiments using cultured bovine urothelial cells showed that this protein is synthesized as a 32-kD precursor that is processed through a 30-kD intermediate, to the mature 27-kD form. In cytoplasmic vesicles containing immature AUM, the AE31 epitope is detected in patches on the cytoplasmic side, but in mature, apical AUM it is detected exclusively on the luminal side. This suggests an unusual translocation of the AE31 epitope during AUM maturation; more data are required, however, to substantiate this interpretation. Immunoaffinity purification of the 27-kD protein results in the copurification in approximately molar ratio of a 15-kD protein, as well as a small and variable amount of a 47-kD protein. Immunoblotting data indicate that these three proteins are immunologically distinguishable. This copurified 15-kD protein is relative basic (pI approximately 8.0). Like the 27-kD protein, it is urothelium-specific and is present mainly in the umbrella cells. Together, our data indicate that a 27-kD protein is urothelial plaque-associated (uroplakin I). Based on complex formation data, we provisionally name the 15-kD protein uroplakin II; additional data will be required to determine whether this and the 47-kD protein are integral parts of AUM. The identification of these AUM-associated and -related proteins, plus the availability of a culture system capable of synthesizing and processing some of these molecules, offer new opportunities for studying the detailed structure, assembly, and function of asymmetrical unit membrane.

Animals↗

Shock, transfusion, and pneumonectomy. Death is due to right heart failure and increased pulmonary vascular resistance.

To determine the physiologic cardiopulmonary abnormalities leading to death when pneumonectomy is required to stop bleeding in patients in hemorrhagic shock, we compared cardiopulmonary responses to resuscitation in pigs undergoing hemorrhagic shock alone, pneumonectomy alone, and hemorrhagic shock plus pneumonectomy. Four shock-plus-pneumonectomy pigs died acutely from right heart failure. When the five remaining shock-plus-pneumonectomy pigs were compared to the two control groups, pulmonary vascular resistance (PVR) increased to significantly higher levels than would be expected from the increase in PVR noted with resuscitation from shock alone and pneumonectomy alone. Right ventricular compensation maintained cardiac index in the hemorrhage-alone group and the pneumonectomy-alone group but could not maintain cardiac index in the shock-plus-pneumonectomy group, despite maximal increases in right ventricular systolic pressure, heart rate, and right ventricular end diastolic volume. These data indicated that resuscitation from shock plus pneumonectomy cannot be effectively accomplished because increased PVR leads to right ventricular failure, which limits left ventricular preload to levels that are insufficient to maintain cardiac index.

Animals↗

Brain mechanisms underlying motor skill learning in the rat.

Young rats subjected to bilateral brain lesions were subsequently tested for acquisition of eight puzzle-box problems. Lesions to 12 of the 37 brain sites investigated (anterior pretectal area, subthalamus, posterolateral hypothalamus, frontocingulate cortex, anterior thalamus, mediodorsal thalamus, ventromedial thalamus, parafascicular nucleus, mamillary bodies, cerebellum, olfactory bulb, and ventromedial hypothalamus) retarded puzzle-box learning. This study, combined with earlier investigation of 11 additional brain sites, show that 21 of the 48 total brain structures are involved in motor skill learning. These findings suggest that motor skill learning requires the integrity of a nonspecific mechanism and several specific mechanisms, such as vestibular-proprioceptive-kinesthetic discrimination mechanism and probably also response flexibility, visuospatial attentional, visual discrimination and place learning mechanisms.

Animals↗

Lung inflation evokes reflex dilation of microvessels in rat skeletal muscle.

Lung inflation can reflexively decrease peripheral resistance, but effects on the microcirculation have not been determined. In this study, we examined the effects of lung inflation on microvessels in skeletal muscle. In anesthetized spontaneously breathing rats, the right cremaster muscle with intact circulation and innervation was exposed and suspended in a tissue bath filled with a physiological salt solution. Diameters of third-order arterioles (3As) were displayed by television microscopy and measured as an index of microcirculatory resistance. In other experiments, sympathetic activity (in the genital femoral nerve) to the cremaster was recorded. Lung inflation decreased systemic blood pressure by 32 +/- 3 mmHg and increased 3A diameter by approximately 23% (P less than 0.05, n = 17). The increase was abolished by cutting the cervical vagus nerves or the right genital femoral nerve. Furthermore, inflation decreased sympathetic nerve activity by 47.6 +/- 11.3% (P less than 0.05, n = 5) before vagotomy but not after. Dilation in response to inflation was also abolished by phentolamine or guanethidine. These results suggest that stimulation of vagal afferents by inflation reflexively decreases microvascular resistance by withdrawal of sympathetic tone.

Animals↗

Production of generalized learning deficit and permanent growth stunting by bilateral brain stem lesions.

Bilateral lesions of the globus pallidus, ventrolateral thalamus, substantia nigra, or the median raphe produce a generalized learning deficit in rats. Bilateral lesions of the dorsomedial hypothalamic nuclei stunt growth in rats without significantly disturbing endocrine functions and without producing a generalized learning deficit. Globus pallidus, ventrolateral thalamus, substantia nigra, median raphe, and dorsomedial hypothalamic nuclei lesions were produced in weanling Sprague-Dawley rats to compare their effect on physical growth. At approximately 72 d of age, all lesions had resulted in reduced body wt, tail length, and tibial length. The differences lacked significance only in body wt after median raphe lesions and tail length after ventrolateral thalamus lesions. In rats with the generalized learning deficit, body size was most stunted after substantia nigra lesions. Tibial epiphyseal width was modestly increased in rats with the generalized learning deficit. Food intake/average body wt ratio in substantia nigra and dorsomedial hypothalamic nuclei rats did not differ significantly from control values. Decreases in brain, heart, liver, kidney, and testes tended to occur after all the lesions, but brain and testis organ wt/body wt ratios were either increased or unchanged. We conclude that brain lesions producing a generalized learning deficit in rats result in impaired physical growth. The results indicated that the stunted animals maintain adequate food intake and have normal growth hormone function. The anatomical substrate for generalized learning impairment may overlap with that of a set point for body size.

Animals↗

Assessing the differentiation state of cultured bovine urothelial cells: elevated synthesis of stratification-related K5 and K6 keratins and persistent expression of uroplakin I.

Although significant progress has recently been made in culturing mammalian urothelial cells, relatively little is known about their biochemical differentiation. In this paper, we assessed the differentiation state of cultured bovine urothelial cells by analyzing their keratins and a cell surface marker, uroplakin I. Urothelial cells were serially cultured either in a serum-free medium, or in a serum-containing medium in the presence of 3T3 feeder cells, with similar results. Despite their stratified appearance, both normal urothelium and cultured urothelial cells synthesize mainly K8, K18 and K19, keratins that are typically seen in simple epithelia. However, cultured urothelial cells synthesize a greatly increased amount of K5 and K6 keratins, which are usually expressed by stratified epithelia but present only in trace amounts in normal urothelium. These data indicate that, as far as keratin synthesis is concerned, cultured urothelial cells undergo an altered pattern of differentiation towards a more 'stratified phenotype'; this unusual finding has interesting implications for urothelial evolution. In the meantime, many superficial cells in cultured urothelial colonies make uroplakin I, a 27 x 10(3) Mr protein subunit of the asymmetrical unit membrane (AUM) characteristic of urothelial (superficial) umbrella cells. These results indicate that cultured urothelial cells undergo, at least in part, AUM biogenesis. Cultured urothelial cells thus provide a useful experimental model system for studying certain early steps of AUM formation.

Animals↗

Local steroid injection: its effect on the recovery of nerve conduction in experimental neuropathy.

This study was designed to investigate the effectiveness of local steroid injection on the recovery of nerve conduction in acute compression neuropathy. Experimental neuropathy with complete conduction block was produced by mechanical compression of the caudal nerves of 24 rats. Five days after compression, the compression site was injected with 0.2 mL Dexamethasone acetate (8 mg/mL suspension) in eight rats (group A), and 0.2 mL normal saline in another eight rats (group B). The remaining eight rats (group C) received no injection. Motor nerve conduction velocity (NCV) over the compressed segment, as well as amplitude of the evoked compound muscle action potential (CMAP) recorded from the segmental tail muscles, were measured before nerve compression and three times per week after nerve compression. The recovery rates of NCV of the caudal nerve were not significantly different among the three groups. The recovery rate of amplitude of CMAP in group A was significantly higher than that in groups B or C (p less than 0.01). However, there was no significant difference in recovery rate of the amplitude of CMAP between groups B and C. It is concluded that local steroid injection to the site of nerve compression may facilitate the recovery of nerve conduction block but not the recovery of demyelinative lesion.

Action Potentials↗

Induction of erythroid differentiation in K562 cells by inhibitors of inosine monophosphate dehydrogenase.

The effects of three inhibitors of inosine monophosphate (IMP) dehydrogenase on a human erythroleukemic cell line, K562, were studied. Following incubation with these inhibitors, K562 cells underwent differentiation and accumulated hemoglobins. The induction of hemoglobin accumulation was dose dependent; maximum induction was observed at 100, 25, and 3 microM, respectively, for ribavirin, tiazofurin, and mycophenolic acid. The induction was associated with reduction of intracellular GTP content and was blocked by adding guanosine within 24 h after adding inducer. The effective dose for half-maximum induction by ribavirin was 3 times less than that for 50% inhibition of K562 proliferation; however, for tiazofurin and mycophenolic acid, it closely approximated the concentrations which suppressed cellular proliferation. Ribavirin was sequestered preferentially inside the K562 cells, and the induction by ribavirin had a greater than 30-fold increase in hemoglobin. Studies with isoelectric focusing, globin chain analyses, and immunochemical assays indicated that both A gamma and G gamma were detected and that the hemoglobin produced in the ribavirin-treated cells consisted of approximately 60% fetal hemoglobin and its acetylated equivalents. The adult-type alpha globin was found, while no beta globin chains were demonstrated. Thus, accumulation of fetal hemoglobin and production of alpha globin chain in ribavirin-treated cells are different from the pattern of hemoglobins induced by hemin.

Cell Compartmentation↗

Transient patterns of acetylcholinesterase activity in developing thalamus: a comparative study in rodents.

This paper describes acetylcholinesterase (AChE) activity in the dorsal thalamus of several rodents, including rat, mouse, gerbil, hamster and guinea pig. Tissue from fetal, neonatal, and adult animals was studied using histochemical techniques. Developing animals of all species display prominent AChE staining in the ventral medial geniculate nucleus and the ventral posterior nucleus, while adult animals show very light staining in these thalamic regions. Similarly, the dorsal lateral geniculate nucleus of all species shows stronger staining in developing animals than in adults. These data indicate that transient expression of AChE activity in primary sensory thalamic nuclei is a characteristic common to a variety of rodents.

Acetylcholinesterase↗