Biomedical subjects

W C Shen

Publications and source records attributed to W C Shen.

At least 73 records · Page 4Linked to original sources

PubMed1994

Transepithelial transport of tyramine across filter-grown MDCK cells via a poly(D-lysine) carrier.

In order to investigate the advantage of using membrane-adsorptive carriers to mediate drug transport across epithelial tissue, we have prepared disulfide- and thioether-linked conjugates of tyramine (tyn) as a model drug to a cationic, nondegradable carrier, poly(D-lysine) (PDL). The transport properties were evaluated using microporous filter-grown Madin-Darby canine kidney (MDCK, strain I) epithelial cells, and we have determined that: (a) the [125I]tyn-SS-PDL conjugate predominantly transported [125I]tyn in the apical-to-basal direction (20-fold greater transport vs. basal-to-apical); (b) [125I]tyn-SS-PDL elicits a 10-fold greater degree of [125I]tyn transport than [125I]tyn-S-PDL, thus demonstrating the importance of the reducible disulfide linkage for [125I]tyn transport to occur; (c) 125I-radioactivity recovered in the basal media was determined to be 95% [125I]tyn-containing small molecules, thus demonstrating the release of [125I]tyn from its PDL carrier; (d) the apical addition of an anionic species, heparin, completely blocks apical-to-basal transport of [125I]tyn, indicating the importance of PDL-mediated binding to the apical membrane for transport to occur; (e) apical-to-basal transport proceeds via non-lysosomal pathways, as lysosomal inactivation by NH4Cl exposure does not inhibit transport, and (f) the addition of a membrane-impermeable inhibitor of disulfide reduction, bisdithionitrobenzoic acid (DTNB), to the apical media inhibits transport by approximately 70%, indicating the importance of apically-localized disulfide reducing reactions for transport of [125I]tyn. Pulse-chase studies indicate that there is no paracellular leakage of conjugate, and the ratio of recycled:membrane-associated:transported [125I]tyn fragment following chase is 4:2:1.

Animals↗

PubMed1994

Subcellular locations of MOD5 proteins: mapping of sequences sufficient for targeting to mitochondria and demonstration that mitochondrial and nuclear isoforms commingle in the cytosol.

MOD5, a gene responsible for the modification of A37 to isopentenyl A37 of both cytosolic and mitochondrial tRNAs, encodes two isozymes. Initiation of translation at the first AUG of the MOD5 open reading frame generates delta 2-isopentenyl pyrophosphate:tRNA isopentanyl transferase I (IPPT-I), which is located predominantly, but not exclusively, in the mitochondria. Initiation of translation at a second AUG generates IPPT-II, which modifies cytoplasmic tRNA. IPPT-II is unable to target to mitochondria. The N-terminal sequence present in IPPT-I and absent in IPPT-II is therefore necessary for mitochondrial targeting. In these studies, we fused MOD5 sequences encoding N-terminal regions to genes encoding passenger proteins, pseudomature COXIV and dihydrofolate reductase, and studied the ability of these chimeric proteins to be imported into mitochondria both in vivo and in vitro. We found that the sequences necessary for mitochondrial import, amino acids 1 to 11, are not sufficient for efficient mitochondrial targeting and that at least some of the amino acids shared by IPPT-I and IPPT-II comprise part of the mitochondrial targeting information. We used indirect immunofluorescence and cell fractionation to locate the MOD5 isozymes in yeast. IPPT-I was found in two subcellular compartments: mitochondria and the cytosol. We also found that IPPT-II had two subcellular locations: nuclei and the cytosol. The nuclear location of this protein is surprising because the A37-->isopentenyl A37 modification had been predicted to occur in the cytoplasm. MOD5 is one of the first genes reported to encode isozymes found in three subcellular compartments.

Alkyl and Aryl Transferases↗

PubMed1994

The establishment of polarity and enhanced transcytosis of transferrin receptors in enterocyte-like Caco-2 cells.

The effect of enterocyte-like differentiation on the transferrin receptor (TfR) polarity in filter-grown Caco-2 cells was studied. The ratio of apical to basolateral TfRs which was found to be approximately 1:1 on the first day after the cells had reached confluence, changed to 1:40 eight days after reaching confluence. The transepithelial electrical resistance (TEER), transport of horseradish peroxidase (HRP) across the monolayer, and total cellular TfR number remained constant over this period. However, the activity of brush border membrane-associated alkaline phosphatase, an established marker for enterocyte differentiation, increased over this 8-day period concurrent with a decrease in apical TfR number. These results suggest that enterocyte-like differentiation rather than tight junction formation is most likely responsible for the polarized distribution of TfRs in Caco-2 cells. The effects of the fungal metabolite brefeldin A (BFA) on TfR distribution and TfR-mediated transcytosis in Caco-2 cells were also studied. BFA caused a marked decrease in the number of basolateral TfRs along with a slight increase in the number of apical TfR. BFA enhanced the TfR-mediated transcytosis of both 125I-Tf and the horseradish peroxidase-Tf conjugate across Caco-2 cells in both apical-to-basolateral and basolateral-to-apical directions. These findings imply a potential application of BFA as an enhancer for TfR-mediated delivery of protein drugs across the intestinal epithelium.

Alkaline Phosphatase↗

PubMed1994

CT of sebaceous adenoma of the parotid gland.

We report a large sebaceous adenoma of the parotid gland. CT showed soft-tissue mass in the lateral portion of the adenoma corresponding to the histologic finding of proliferation of sebaceous glands and fibrous stromal tissue. There were numerous fatty cysts (-65Hu). Some of the cystic component ruptured, and CT demonstrated leaking of fatty material into the parapharyngeal space.

Adenoma↗

PubMed1994

Percutaneous transluminal angioplasty of neck vessels.

BACKGROUND: Although surgical endarterectomy or bypass is regarded as the treatment-of-choice for extracranial cerebral vascular stenosis, percutaneous transluminal angioplasty (PTA) has its role as an alternative procedure for symptomatic patients refractory to medical therapy and/or for whom surgical treatment is contraindicated. METHODS: Occlusive blood vessels were first identified by neurologic, ultrasonic and angiographic studies. Patients were premedicated with heparin and corticosteroid. From a transfemoral approach, double lumen balloon dilatation catheter was guided across the area of stenosis, and inflated two or three times to produce dilatation. Following PTA, patients received anticoagulants and corticosteroid for a period, and were followed by clinical examination, Doppler blood-flow and angiographic studies. RESULTS: Seventeen patients undergoing 26 procedures were included in this study. The lesions involved subclavian artery in nine cases, common carotid artery in seven, internal carotid artery in five and proximal vertebral artery in five. Digital subtraction angiography (DSA) was done immediately after PTA and showed improvement of stenotic segment (residual stenosis < 30%) in 22 vessels. Restenosis occurred in two cases of subclavian artery PTA six months later. There were three complications, including TIA in two cases and hematoma at the puncture site in one case. Clinically, symptomatic improvement was obtained in 13 cases. CONCLUSIONS: In patients with significant atherosclerotic stenosis of neck vessels, PTA is a beneficial procedure for improving blood flow to brain circulation and for alleviating symptoms.

Aged↗

PubMed1994

MR imaging evaluation of bone marrow signal change in post-irradiation patients with nasopharyngeal carcinoma.

BACKGROUND: The normal distribution of red and yellow marrow can be altered by therapeutic irradiation, which causes a well-documented sequence of bone marrow changes. This study measured the T1 signal intensity of the clivus and cervical vertebral bodies of NPC patients who had received a complete course of radiation therapy. METHODS: There were 3 categories of patients: Group I: NPC patients who had radiation myelitis; 14 persons which a total 20 times of MRI. Group II: NPC patients, without radiation myelitis; 6 patients, which a total 6 times of MRI. Group III (control group): patients who received MRI of the cervical spine because of cervical spondylosis or HIVD, were total of 45 patients. In the sagittal section of T1WI, the signal intensity of the bone marrow of clivus and C2 to T1 vertebral bodies was measured. RESULTS: 1). There were homogeneous increases of signal intensity of the bone marrow of clivus and C2 to T1 in Groups I and II. 2). There was no statistical difference between Groups I and II. 3). The increased signal intensity of bone marrow after radiation showed no difference in short and long duration between radiation therapy and MR examinations, indicating that bone marrow signal intensities were increased in NPC patients whether or not they had radiation myelitis. CONCLUSIONS: Signal change in bone marrow may have occurred soon after radiation therapy, and may have persisted for several years. The radiation myelitis always involved the low medulla oblongata to C5 level; however, the bone marrow signal change always extended downward to the T1 level, so bone marrow is more sensitive than the spinal cord and is prone to be affected by irradiation.

Adult↗

PubMed1993

The Saccharomyces cerevisiae LOS1 gene involved in pre-tRNA splicing encodes a nuclear protein that behaves as a component of the nuclear matrix.

Mutations of the Saccharomyces cerevisiae LOS1 gene cause the accumulation of end matured intron-containing pre-tRNAs at elevated temperatures. In an effort to decipher the role of the LOS1 protein in pre-tRNA splicing, we have analyzed the LOS1 gene and its protein product. The LOS1 gene is located on the left arm of chromosome XI and the order of genes in this area of the chromosome is .... URA1 ... SAC1 TRP3 UBA1 STE6 LOS1 .... FAS1..... The LOS1 open reading frame encodes a putative protein of 1100 amino acids that shows no significant homology to other genes. The LOS1 open reading frame was tagged with the influenza virus hemagglutinin epitope recognized by the 12CA5 antibody. The 12CA5 antibody recognizes an epitope-tagged protein of the size predicted by the LOS1 open reading frame. Using this antibody for indirect immunofluorescence and cell fractionation studies we show that the LOS1 protein is located in nuclei. Los1p cannot be extracted from nuclei by treatment with nucleases, salts, or Triton X-100. This insolubility suggests that Los1p is a component of the nucleoskeleton. We propose that LOS1 mutations may affect pre-tRNA processing via alteration of the nuclear matrix.

Amino Acid Sequence↗

PubMed1993

Disseminated tuberculomas in spinal cord and brain demonstrated by MRI with gadolinium-DTPA.

Intramedullary tuberculoma is rare, and there has been no report of concurrent intramedullary and intracerebral tuberculomas. We report a 30-year-old man with miliary tuberculosis of the lung. He suffered sudden paraplegia due to tuberculomas in the thoracic spinal cord and MRI showed more tuberculomas in the cervical spinal cord, brain stem, and cerebral and cerebellar hemispheres. The tuberculomas were isointense on the T1-weighted images, and hyperintense on the T2-weighted images; there was marked enhancement with intravenous gadolinium-DTPA. All the tuberculomas were very small 1 year after antituberculous chemotherapy.

Adult↗

PubMed1993

Myelography, CT and MRI in leukaemic infiltration of the lumbar theca.

A 25-year-old woman with acute lymphoblastic leukaemia, while in remission, developed paraparesis, with faecal and urinary incontinence. CT demonstrated increased density of the lumbar theca and enlargement of the nerve roots. Myelography showed complete obstruction below the L3 level. MRI showed increased signal intensity in the lumbar sac on T1 weighting, and the cauda equina enhanced with gadolinium-DTPA. Lymphoblasts were seen in the lumbar spinal fluid. After chemotherapy, these abnormalities resolved, as did the paraparesis and incontinence.

Adult↗

PubMed1993

Wallenberg's lateral medullary syndrome with loss of pain and temperature sensation on the contralateral face: clinical, MRI and electrophysiological studies.

Thirteen patients with Wallenberg's lateral medullary syndrome (WLMS) were studied. Clinical and magnetic resonance imaging (MRI) evidence demonstrated infarction in the dorsolateral medulla which produced loss of pain and temperature sensation on one side of the face ipsilateral to the lesion in seven patients. However, in another six patients, the infarction in a similar location produced the same sensory loss on the contralateral face. This is the first report of an analysis comparing these two conditions in WLMS patients, confirmed by MRI. The finding of normal blink reflex and somatosensory evoked potentials (stimulation on median nerve) in the two groups of patients may indicate that the impulses travel along the central pathways of touch, vibration and joint position sensation instead of the pathways for pain and temperature, because the patients had normal sensation of touch, vibration and joint position but impairment of pain and temperature sensation. In addition, it is suggested that the pathways of late blink reflex (R2) pass through the medial lemniscus in the ventromedial medulla instead of the spinal trigeminal tract in the dorsolateral medulla. Further, the observation of the much longer lantencies (about 29 ms) of the normal R2 raises the possibility that the impulses may travel along the longer pathways through the opposite medial lemniscus and up to the thalamus or cortex where they project to bilateral motoneurons of the orbicularis oculi muscles. Although the alternative of R2 travelling only by the shorter pathways through the brain stem is not excluded, this is not supported by current data.

Adult↗

PubMed1993

Regulation of pathways within cultured epithelial cells for the transcytosis of a basal membrane-bound peroxidase-polylysine conjugate.

A conjugate of horseradish peroxidase (HRP) to poly(L-lysine) (PLL) was used as a non-specific adsorptive probe to study transcytosis in MDCK strain I and Caco-2 epithelial cells. As we have shown previously, HRP-PLL transcytosis proceeds via an intracellular, non-lysosomal proteolytic compartment in MDCK cells; yet, this compartment is utilized for transcytosis only in the basal-to-apical direction (Taub, M. E. and Shen, W.-C. J. Cell. Physiol., 150, 283-290, 1992). Using size exclusion chromatography, we demonstrate that the PLL moiety of the conjugate is effectively cleaved during transcytosis, thus releasing free HRP from the apical surface of the cells. Pulse-chase studies indicate that approximately 6% of basolateral surface-associated HRP-PLL conjugate in Transwell-grown cell monolayers enters the basal-to-apical transcytotic pathway. Brief (1 hour) treatment with 160 nM phorbol ester (PMA), a protein kinase C stimulator, elicits a 2-fold increase in the rate and amount of HRP-PLL transcytosis following a 1 hour lag time. Treatment with 1.6 micrograms/ml brefeldin A (BFA) inhibits HRP-PLL transcytosis by approximately 30%; additionally, BFA is able to abolish completely the PMA stimulatory effect. Removal of BFA causes a re-establishment of the normal rate of transcytosis within 2 hours, demonstrating the reversibility of BFA inhibition with respect to HRP-PLL transcytosis. Thus, PMA most likely elicits an increase in the amount of basally internalized conjugate delivered to BFA-sensitive transcytotic compartments.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Cationized Bowman-Birk protease inhibitor as a targeted cancer chemopreventive agent.

The conjugate of the Bowman-Birk inhibitor (BBI) with poly(D-lysine) (PDL-ss-BBI) has been suggested as a lung-targeted anti-carcinogenic agent. The authors demonstrate that PDL-ss-BBI, given i.p., reduces the tumor number in the lungs of 3-methylcholanthrene treated mice (61-71% compared to control group) in a dose-dependent manner, but is toxic to the treated animals at a high dosage. In order to develop a better lung-targeted anti-carcinogenic agent, spermine-conjugated BBI (spermine-BBI) was synthesized by coupling BBI to spermine through amide bonds using a carbodiimide-mediated reaction. Results from in vitro transformation assays demonstrated that spermine-BBI was at least as effective as BBI in reducing the transformation yield in C3H10T1/2 cells. When injected intravenously into mice [125I]spermine-BBI accumulated to a greater extent in the lungs and the liver compared to BBI. The in vitro cytotoxicity of spermine-BBI in C3H10T1/2 cells was 30-fold less than that of PDL-ss-BBI. These results suggest that spermine-BBI is likely to be an improved cancer chemopreventive agent compared to BBI or PDL-ss-BBI.

Animals↗

PubMed1993

Magnetic resonance imaging of cerebral hemiatrophy.

Twenty-one cases of cerebral hemiatrophy were studied using 1.0 T magnetic resonance imaging (MRI). In addition to showing the generalized decrease in volume of the cerebral hemisphere, three patterns of cerebral hemiatrophy were noted: pattern I, diffuse cortical and subcortical atrophy (six cases):pattern II, diffuse cortical atrophy associated with expanded porencephalic cysts (10 cases); pattern III, old infarction with necrosis in the territory of the middle cerebral artery (five cases). Pattern I and pattern II were believed to have resulted from hypoxic-ischemic encephalopathy with subcortical leukomalacia (SCL). In all 21 cases, MRI also showed Wallerian degeneration, hyperintensive signals in the posterior limb of the internal capsule on the T2-weighted images, and unilateral atrophy of the cerebral peduncle as far as the pyramid of the medulla oblongata. Structural changes such as thickness of the calvarium, dilatation of the ipsilateral side of the frontal and ethmoid sinus, and reduced size of the middle cranial fossa, were found in 13 cases. Thus, MRI is a useful method of examination in the diagnosis of cerebral hemiatrophy as it has the ability to highlight changes in the cerebral hemisphere, basal ganglion, thalamus and brainstem as well as highlighting bony structural changes.

Adolescent↗

PubMed1993

[MRI of postmortem brains].

We studied 18 formalin-fixed brains using MRI, and correlated our data with subsequent gross and microscopic examinations. 9 of our patients died from brain diseases (stroke due to infarction 4, stroke due to hemorrhage 1, encephalitis 2, head injury 1, brain tumor 1). 9 of our patients died from non-CNS diseases (stomach cancer 1, colon cancer 1, liver cirrhosis 1, myocardial infarction 2, trauma 4). In MRI of postmortem brain, T1WI and T 2WI was able to clearly show the myelination process of brainstem, basal ganglia, internal capsule and optic radiation in a 2 months-old-boy. The findings were similar to MRI of live infants. In normal adult postmortem brains, the T1WI showed a relatively low signal intensity of white matter as compared to gray matter. The pictures were similar to proton density images, not T1WI of normal adult brains. The reason why the signal intensity of the white matter was lower than the gray matter may have been due to lysis of lipid of myelin sheath in the formalin solution. Postmortem MRI was able to detect the periventricular hyperintensity (corresponding to arteriosclerotic encephalopathy) and subcortical hyperintensity spots (which corresponding to the widening of the Virchow-Robin perivascular space because of arteriosclerosis) in the brains of our elderly patients. Postmortem MRI detected the intracerebral hemorrhage, which appeared as a dark signal in both short and long TR images. However, MRI did not show blood in the ventricles, sulci, or superficial hemorrhages in the cortex of brain. Brain edema was revealed in the postmortem MRI and appeared as low signal intensity in T1WI and hyperintensity in T2WI. It was associated with a significant mass effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

PubMed1993

[The chemical shift imaging of liver].

Tissue characterization of different pathologies of the liver can be achieved by differences of relaxation time on changing of pulse sequences in magnetic resonance imaging (MRI). The usefulness of MRI for detection of liver disease is limited when the pathological change is subtle. Chemical shift is a fundamental characteristics in nuclear magnetic resonance (NMR). Chemical shift imaging (CSI) in MRI is based on the different Larmor frequency of proton in water and fat; and therefore it was able to enhance the effectiveness of pathology. For this study, Dixon's method was used to detect liver pathologies and compare its detectability with conventional pulse sequences. Forty cases were enrolled for study; they included 5 health volunteers, 15 hepatomas, 1 cholangiocarcinoma, 5 metastatic hepatic tumors and 14 fatty livers. In hepatic tumors, the lesion number, tumor margination and lesion-to-liver contrast in images were read and analyzed. Signal intensities, signal-to-noise ratio and contrast-to-noise ratio were compared, after measurement, from stored data. In fatty livers, the relative change of signal intensities in different areas of the liver in in-phase and out-phase images were compared with the back muscle and spleen to find where the fatty metamorphosis happened. CSI in spin echo or gradient echo pulse sequences was found to be adequate and valuable for detecting fatty liver, when compared with conventional MRI. CSI not only identified the extension of disease itself but also characterized the fatty change in liver parenchyma. Though CSI affords no further advantages than conventional pulse sequences for detection of hepatic tumors, occasionally, when the image quality of the conventional pulse sequences is not satisfactory or equivocal in lesion detection, the use of CSI might be attempted.

Fatty Liver↗

PubMed1993

A clinical and MRI study of cerebellar infarctions.

The frequency of cerebellar infarctions over two and a half years was 2.7% of the 1300 hospitalized patients over that period. Sixteen patients with cerebellar infarctions were studied by using clinical manifestations and magnetic resonance imaging (MRI). Ages ranged from 41-87 (mean 63.5) years; 13 were men and 3, women. Risk factors included: hypertension (50%), diabetes (44%), prior stroke (44%), cardiac disease (38%), and hyperlipidemia (19%). Common symptoms and signs were dizziness/vertigo (75%), unsteadiness (69%), dysarthria (69%), and nausea/vomiting (50%). Infarcts mainly involved the posterior inferior cerebellar artery (PICA) territory and tended to be associated with brainstem infarcts in 14 of the 16 patients. Most cerebellar infarctions had a benign course, especially the small ones. No mortality was noted in this series. The short-term outcome of the cerebellar infarctions seemed to depend on the size of the infarcts and the sites of the artery occlusion. It was concluded that diagnosis of cerebellar infarctions requires a high index of clinical suspicion, especially when patients present with a sudden onset of ataxia, dizziness/vertigo, nausea/vomiting and dysarthria; and that MRI is a useful tool for the detailed study of cerebellar infarctions and can elucidate associated brainstem infarcts.

Adult↗

PubMed1993

Gadolinium-DTPA enhanced magnetic resonance imaging of dural sinus occlusion in dural arteriovenous malformation: a case report.

A case of dural arteriovenous malformation of the transverse sinus, is presented, with dural sinus occlusion imaged with gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA). The patient, a 59-year-old man, had developed easy forgetfulness and frequent misreading over a four-month period; he had had episodic vertiginous attacks since 3 January 1992. Magnetic resonance (MR) imaging done on 21 January 1992 showed abnormally dilated subcortical vessels and a non-hemorrhagic venous ischemia in the subcortical white matter of the left posterior temporal lobe. Thrombosis of the left sigmoid sinus was seen. After administration of intravenous Gd-DTPA, dense enhancement in the left sigmoid sinus and abnormal serpiginous enhancement in the left temporal lobe were revealed. Left carotid angiography showed dural arteriovenous malformation in the posterior fossa, with non-opacification of the left transverse and sigmoid sinuses, plus forward venous drainage into left superior petrosal sinus. The focal neurologic deficits in this case from non-hemorrhagic venous ischemia in the subcortical white matter of the left posterior temporal lobe, and were secondary to venous hypertension.

Dura Mater↗

PubMed1993

Detection of renal allograft dysfunction with color Doppler flow ultrasound.

Thirty-six color Doppler flow (CDF) ultrasound studies were performed on 28 allograft kidneys to assess the capability of this technique in diagnosing transplant dysfunction through morphology and Doppler spectra. The composite score obtained from ultrasound images showed no statistical significance. Quantitative studies with Pulsatility Index (PI) and Resistive Index (RI) after localization of the arcuate, interlobar, segmental and main renal arteries by CDF revealed significant differences between normal functioning graft kidneys and the group of acute rejection, as well as the group of acute tubular necrosis (ATN) (P < 0.05). No apparent change was demonstrated in patients with chronic rejection. When PI was greater than 1.60 or RI was greater than 0.80, the transplant kidney was more likely to be rejected, though possibility of ATN should be considered. RI was easier to calculate, and correlated better than PI with the serum creatinine level. CDF expedited the Doppler spectrum study.

Adult↗