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Biomedical subjects

J M Lee

Publications and source records attributed to J M Lee.

At least 127 records · Page 7Linked to original sources
PubMed2000

Vitellogenin of the cicada Graptopsaltria nigrofuscata (Homoptera): analysis of its primary structure.

We cloned and sequenced the cDNA of vitellogenin (Vg) from the cicada Graptopsaltria nigrofuscata (Homoptera). The deduced amino acid sequence of 1987 residues (including 16 residues for a putative signal peptide) was obtained. The pro-Vg was cleaved into two subunits between residues 379 and 380 following a consensus RXXR cleavage site sequence, secreted as S-Vg (apparent molecular weight 43 kDa) and L-Vg (200 kDa), sequestered, and stored in the egg as two vitellins (Vns), S-Vn and L-Vn, with similar respective molecular weights. There was a single long serine-rich stretch closely following the cleavage site. The entire amino acid sequences of the Vgs from the eight insects so far reported could be aligned confidently. The presence of subdomains I-V (areas of relatively high amino acid conservation) and of 10 cysteines at conserved locations at the C-terminus, noted previously among insect Vgs, were confirmed. Antisera raised against G. nigrofuscata S- and L-Vn cross-reacted with the S- and L-Vg/Vn, respectively, of all three other cicada species examined. Another major egg protein (170 kDa) unrelated to Vg/Vn, was also detected in all species examined.

Amino Acid Sequence↗
PubMed2000

A simple and rapid method for cloning insect vitellogenin cDNAs.

We describe a simple and rapid method for cloning insect vitellogenin (Vg) cDNAs. The method relies on the facts that insect Vg amino acid sequences can be aligned confidently along their entire lengths and that a short, highly conserved GL/ICG motif and up to nine cysteine residues that follow at conserved locations are present near the C-termini. An adaptor-ligated double-strand cDNA library is constructed from poly(A)+ RNA prepared from vitellogenic female fat body tissues using a commercial kit, and subjected to PCR with each of the degenerate nucleotide sequences for the GL/ICG motif and the adaptor sequence as primers. The PCR products (0.7-0.9 kb, representing the 3' portion) are cloned, the nucleotide sequences are determined, and the deduced amino acid sequences are aligned with the known insect Vg sequences starting from the GL/ICG motif. Gene-specific primers corresponding to the sequences near the 5'-termini of the initial clones and the adaptor sequence are employed to obtain the remaining 5' portion of the Vg cDNAs. The method was successfully applied to the bean bug Plautia stali (Heteroptera), revealing three Vg genes.

Amino Acid Sequence↗
PubMed2000

Conservation of the elastic and flexural moduli of osteopenic femoral cortical bone in experimental inflammatory arthritis in the rabbit.

Experimental inflammatory arthritis (EIA) produced by carrageenan injection provokes a rapid bone remodeling state with cortical and cancellous bone loss. The objective of this study was to determine whether changes in cortical mechanical properties and/or geometry occur in long bones, either near or remote to the site of inflammation. EIA was induced in the right tibio-femoral joint of rabbits over 56 days. The right humerus and right femur from 15 normal and 25 arthritis group animals were excized. Semi-cylindrical specimens of the medial cortical shaft were subjected to non-destructive four-point bending tests. Transverse sections at the four contact sites of the loading jig were photographed and digitized to obtain average cross-sectional area (A) and moment of inertia (I). Moment of inertia and slope of the load/deflection curve permitted calculation of modulus of elasticity (E) for each specimen. Load/time curves were also used to calculate per cent stress remaining in relaxation experiments. Per cent stress remaining, E, A, I and square root of I/A (radius of gyration) were examined for differences by bone (humerus, femur) and by treatment (N,A) using two way ANOVA. The induction of inflammatory arthritis did not significantly alter the modulus of elasticity in either the femur or humerus; however, arthritis reduced the moment of inertia from 34.54 +/- 2.88 x 10(-12) m4 to 25.06+/-1.80 x 10(-12) m4 (mean+/-SEM, p < 0.05). This was observed in the femur (near the arthritic joint), but not in the humerus (remote from arthritic joint). Analysis of area and ratio I/A demonstrated that this geometric effect of treatment was due to reduced area without gross cross-sectional shape changes. Per cent stress remaining in the femur (but not in the humerus) was higher in the arthritis specimens than in the normal specimens (N: 80.86 < 0.97%; A: 83.25 +/- 0.71%, p < 0.05). Thus, in this arthritis model, the principal mechanical or geometric effect on cortical bone was reduction of the cross-sectional area and moment of inertia. The viscoelastic relaxation response of bone was also altered, perhaps due to loss of water or collagen degradation.

Journal Article↗
PubMed2000

AAC-11 overexpression induces invasion and protects cervical cancer cells from apoptosis.

To identify the genes involved in cervical carcinogenesis, we applied the mRNA differential display (DD) method to analyze normal cervical tissue, cervical cancer, metastatic lymph node, and cervical cancer cell line. We cloned a 491-bp cDNA fragment, CC231, which was present in metastatic tissue and cervical cancer cell line, but absent in normal cervical and cervical cancer tissues. The 491 bp cDNA fragment has 98% homology to the previously published sequence, AAC-11 (antiapoptosis clone 11). The levels of AAC-11 mRNA expressions in nine normal cervical and nine primary cervical cancer tissues were low. Its expression was higher in three metastatic tissues and five cervical cancer cell lines (HeLa, CaSki, SiHa, CUMC-3, and CUMC-6). Invasion of matrigel and adhesion to laminin by AAC-11 transfected CUMC-6 cells were increased by approximately 2-fold and 4-fold, respectively. Northern blot analysis showed that matrix metalloproteinase (MMP)-2 and membrane type 1 MMP (MT1-MMP) genes were found to be expressed in high levels in AAC-11-transfected cancer cells. But MMP-2 and MT1-MMP were not expressed in cells transfected with vector alone or wild-type cells. AAC-11-transfected cells expressed an elevated level of MMP-2 protein as assessed by immunoblotting. On the contrary, tissue inhibitor of MMP (TIMP-2) expression was detectable in cells transfected with vector alone or wild-type cells, respectively. Its expression was undetectable in AAC-11 transfected cells. In cervical cancer cells transfected with AAC-11, the expression of beta-catenin was up-regulated. These suggest that overexpressions of MMP-2 and MT1-MMP, loss of TIMP-2 expression, and up-regulation of beta-catenin by AAC-11 transfection may contribute to the development of cervical cancer invasion. AAC-11 gene transfection increased cervical cancer cell colonization. The effect of AAC-11 on cultured cervical cancer cells was associated with antiapoptotic process. Approximately 50% of the AAC-11 transfected cells in serum-free medium died after 2 weeks, compared to 1 week for vector alone or wild-type cells. These results suggest that AAC-11 may serve as a candidate metastasis-related and apoptosis-inhibiting gene in human cervical cancer.

Apoptosis↗
PubMed2000

Microsatellite analysis in post-transplantation lymphoproliferative disorder to determine donor/recipient origin.

Post-transplantation lymphoproliferative disorders (PTLD) are a group of heterogeneous diseases that occur after organ transplantation. Determination of the origin of the tumor cells not only provides clues to its possible pathogenetic mechanism, but also gives prognostic guidance in the clinical management of patients. We reviewed the clinicopathological features of four cases of PTLD that developed after solid organ transplantation. Using microsatellite analysis performed on paraffin-embedded tissue and using multiple, highly polymorphic markers, we have successfully determined the recipient/donor origin of the tumor cells in all of them. The time of onset of the PTLD ranged from 5 to 11 mo. All cases were diffuse large cell lymphomas of B-cell lineage, and the two cases that have been tested for EBV by in situ hybridization were positive. Three of the 4 PTLD were of donor origin and these three patients died of diseases unrelated to PTLD. The single patient with PTLD of recipient origin died of disseminated PTLD. The mean survival length of the three patients with donor origin was 26.3 mo, whereas that of the patient with recipient origin was 12 mo. Our results indicate a relatively high incidence of PTLD of donor origin among our patients with solid organ transplantation, as compared to other reported series. Moreover, the finding of the relatively indolent nature of PTLD of donor origin supports that determination of the donor/recipient origin of PTLD is of prognostic significance.

Adult↗
PubMed2000

Use of esophagectomy to treat recurrent hyperplastic tissue obstruction caused by multiple metallic stent insertion for corrosive stricture.

We report a case of a 75-year-old woman who received repeated metallic stent insertion for corrosive esophageal injury. She underwent esophagectomy and gastric tube reconstruction about 3 years after injury because both stents were occluded in turn by overgrowth of granulation tissue. The gross and microscopic changes of the esophagus secondary to prolonged stent insertion are described. In the literature, no reports of similar cases have been recorded. Our limited experience revealed that using metallic stents to treat benign esophageal stricture should be handled very cautiously because of the complications which can commonly occur and are difficult to manage. Repeated stent insertion, although effective for temporarily relieving dysphagia, is ineffective in the long run and can create complications. We suggest that the feasibility of esophagectomy should be evaluated after the improvement of the general condition of the patient.

Aged↗
PubMed2000

Neuronal apoptosis after CNS injury: the roles of glutamate and calcium.

While a role has been well established for excitotoxic necrosis in the pathogenesis of traumatic or ischemic damage to the CNS, accumulating evidence now suggests that apoptosis may also be a prominent contributor. In this review we focus on the role of glutamate and attendant intracellular calcium influx in triggering or modifying excitotoxic necrosis and apoptosis, raising the possibility that calcium influx may affect these two death pathways in opposite directions. Incorporating consideration of both pathways will probably be needed to develop the most effective neuroprotective treatments for CNS injury.

Animals↗
PubMed2000

Transglutaminase-induced cross-linking of tau proteins in progressive supranuclear palsy.

The mechanisms leading to the abnormal self-polymerization of tau into straight and paired helical filaments (PHFs) and neurofibrillary tangles (NFT) in Alzheimer disease (AD) and progressive supranuclear palsy (PSP) are not known. However, transglutaminase-induced cross-linking of PHF-tau was observed in AD and thus may also contribute to the formation of NFT in other neurodegenerative disorders including PSP. Tissue homogenates from PSP and normal age-matched controls were used to immunoaffinity-purify proteins containing transglutaminase-induced epsilon-(gamma-glutamyl) lysine cross-links. The immunoaffinity-purified proteins were then examined on immunoblots with a PHF-tau antibody, PHF-1. There were significantly higher levels of epsilon-(gamma-glutamyl) lysine cross-linking of PHF-tau in globus pallidus and pons regions of PSP cases compared to barely detectable cross-links in controls. The occipital cortex, an area spared from neurofibrillary pathology in PSP, showed no detectable cross-linking of PHF-tau protein in either PSP cases or control cases. Double-label immunofluorescence demonstrated the colocalization of the cross-link and PHF-tau in NFT in pons of PSP Previous studies and present data are consistent with the hypothesis that transglutaminase-induced cross-linking may be a factor contributing to the abnormal polymerization and stabilization of tau in straight and PHFs leading to neurofibrillary tangle formation in neurodegenerative diseases, including PSP and AD.

Aged↗
PubMed2000

MR characteristics of progressive massive fibrosis.

This study was performed to evaluate the static and dynamic magnetic resonance imaging (MRI) contrast enhancement pattern of progressive massive fibrosis (PMF) in coal workers' pneumoconiosis. Eighteen lesions in 12 patients were evaluated using a 1.5-T MR unit. T1-weighted FLASH images were obtained before and 0.5, 1, 2, 3, 4, 5, 7.5, 10, 12.5, and 15 minutes after injection of gadolinium diethylenetriamine penta-acetic acid (Gd-DTPA; 0.1 mmol/kg). Imaging findings, the contrast enhancing pattern, enhancement time curve, and the contrast uptake equivalent (CE; mmol/L) were evaluated. On T1-weighted images, 14 lesions showed high signal intensity, and four showed low signal intensity. On T2-weighted images, all lesions were of low signal intensity and were indistinguishable from aerated lung parenchyma. After contrast infusion, all lesions except two enhanced markedly. The time enhancement curve showed a marked, gradual increase in signal intensity up to 3 minutes, a subtle increase in signal intensity up to 7.5 minutes, and then a plateau until 15 minutes after Gd-DTPA injection. Characteristic MR findings of PMF in coal workers' pneumoconiosis include T1 high signal intensity, T2 low signal intensity, and marked postinfusion enhancement. The time enhancement curve shows a marked, gradual increase of signal intensity up to 3 minutes and plateau up to 15 minutes.

Aged↗
PubMed2000

Rapid urease tests lack sensitivity in Helicobacter pylori diagnosis when peptic ulcer disease presents with bleeding.

OBJECTIVE: The eradication of Helicobacter pylori (H. pylori) in patients with bleeding peptic ulcer disease (PUD) decreases the rate of ulcer rebleeding. Although all methods for H. pylori diagnosis have been extensively evaluated in uncomplicated PUD the efficacy of the commonly used rapid urease test (RUT) has not been established in patients with bleeding peptic ulcer disease. The aim of this study was to evaluate the efficacy of the RUT (CLOtest) in patients with bleeding duodenal ulcers (DUs). METHODS: Consecutive patients with symptoms of upper GI tract hemorrhage and a DU at the time of endoscopy were evaluated. The presence of H. pylori infection was determined by RUT, microbiology, and histology. Consecutive patients with uncomplicated DUs were similarly evaluated. The prevalence of H. pylori as determined by the RUT alone was compared to that determined by a combination of all tests in both patient groups. RESULTS: Fifty-five patients with bleeding DUs and 69 with nonbleeding DUs were evaluated. The prevalence of H. pylori in patients presenting with bleeding was 72.7% (95% confidence interval [CI] 61.0-84.5%) and lower than the prevalence rate of 92.8% (95% CI 86.6-98.8%) in patients with uncomplicated PUD (p < 0.05). The prevalence of H. pylori in the bleeding DU group as determined by RUT alone (54.5%) was less than that determined by a combination of all tests (73%) with a false-negative rate of 10 of 40 (25%; 95% CI 11.6-38.4%) (p < 0.05). This false-negative rate was significantly greater than that observed in the group presenting with dyspepsia (1 of 64 [1.6%; 95% CI 0-4.6%]) (p < 0.01). CONCLUSIONS: The prevalence of H. pylori is lower in patients with bleeding DUs when compared to patients with uncomplicated DUs. In addition, the sensitivity and negative predictive value of the RUT is lower in patients presenting with bleeding, and other methods of H. pylori diagnosis should be used in this patient group.

Adult↗
PubMed2000

Increased numbers of both airway basophils and mast cells in sputum after allergen inhalation challenge of atopic asthmatics.

Mast cells and basophils are metachromatic cells that participate in allergic inflammation. Allergen challenge to the airways of atopic asthmatic individuals increases levels of metachromatic cells, which may reflect an increase in mast cells, basophils, or both. We conducted a study to characterize the kinetics of basophil and mast cell recruitment to the airways of atopic asthmatic subjects after allergen inhalation challenge, using monoclonal antibodies specific for each type of cell. Of 19 subjects, 14 developed both early- and late-phase asthmatic responses (dual responders [DRs]), whereas five developed only early asthmatic responses (early responders [ERs]) after allergen inhalation. There was a significant increase in the number of sputum eosinophils (p < 0.002) and basophils (p < 0.002) at 7 h and 24 h after challenge in both ERs and DRs. There was also a significant increase in the number of activated eosinophils (p = 0. 00002) and mast cells (p = 0.009) in sputum at 7 h and 24 h after challenge in DRs, but not in ERs (p > 0.4). DRs had a significantly higher number of allergen-induced sputum basophils than did ERs (p < 0.01), and sputum basophils correlated significantly with airway hyperresponsiveness (AHR) to methacholine at 24 h after challenge (r = 0.66, p = 0.002). DRs tended to have higher allergen-induced basophil levels than did ERs, which may contribute to the observed AHR.

Adult↗
PubMed2000

The usefulness of orbital lines in detecting blow-out fracture on plain radiography.

The purpose of this study was to assess the diagnostic value of orbital line changes on plain radiographs in detecting blow-out fracture. 92 cases of surgically confirmed blow-out fracture were retrospectively analysed in regard to plain radiographs and CT. Anterior and posterior lamina papyracea lines (ALPL and PLPL, respectively) of the orbital medial wall as well as the posteromedial floor line (PMFL) were assessed on orbital posteroanterior projections. The inferomedial orbital line (IMOL) and the inferior wall line were assessed on Waters projections. Orbital lines on plain radiographs were compared with CT findings. Of 53 cases of lamina papyracea fracture, 47 cases showed orbital line changes on plain radiographs (sensitivity 88.7%). Changes of PLPL (41/47), IMOL (29/47), ALPL (4/47) and PMFL (2/47) were demonstrated as depression, loss, discontinuity and irregularity. In four cases showing normal radiographs, the fractures measured less than 9 mm in size on CT. All 25 cases with orbital floor fracture accompanying medial wall fracture demonstrated orbital line changes of PMFL (16/25), PLPL (14/25), ALPL (3/25) and IMOL (2/25) (sensitivity 100%). 12 of 14 cases with orbital floor fracture demonstrated changes of inferior wall line (7/12) and PMFL (6/12) (sensitivity 85.7%). Two cases demonstrated asymmetric focal soft tissue density without orbital line changes. Orbital line changes on plain radiographs corresponded well with CT findings, confirming the usefulness of plain radiographs in detecting blow-out fracture.

Adolescent↗
PubMed2000

A linkage map of the turnip sawfly Athalia rosae (Hymenoptera: Symphyta) based on random amplified polymorphic DNAs.

A linkage map was constructed for the sawfly, Athalia rosae (Hymenoptera), based on the segregation of random amplified polymorphic DNA (RAPD) markers and a visible mutation, yellow fat body (yfb). Forty haploid male progeny (20 yfb and 20+) from a single diploid female parent (yfb/+) were examined. Sixty-one of the 180 arbitrary primers tested by polymerase chain reaction (PCR) produced one or more RAPD bands. A total of 79 RAPD markers were detected. Of these, seven showed significant deviation from the expected 1:1 ratio, and were therefore excluded from further analysis. The remaining 72 RAPD markers and the marker mutation, yfb, were subjected to linkage analysis. Sixty RAPD markers and the yfb marker were organized into 16 linkage groups, spanning a distance of 517.2 cM. Twelve RAPD markers showed no linkage relationship to any group. Thirteen gel-purified RAPD bands were cloned and sequenced to generate the sequence-tagged sites (STSs). A single locus was represented by two markers, with one of them having a short internal deletion.

Animals↗
PubMed2000

Biphasic anaphylactic reactions in pediatrics.

OBJECTIVES: The objectives of this study were to: 1) determine the incidence of biphasic reactions in children with anaphylaxis; 2) establish what risk factors can predict progression to a biphasic reaction; and 3) assess the utility of inpatient observation for patients whose anaphylaxis appears to have resolved. METHODS: We performed a retrospective analysis of all children admitted to Children's Hospital inpatient service between 1985 and 1999 with acute anaphylaxis. Data were collected from the medical records regarding past medical history, presenting signs and symptoms, treatment, and hospital course. Patients were considered to have resolution of anaphylaxis if they were documented to have cessation of all symptoms and needed no therapy for at least 1 hour. Biphasic reactions were defined as a worsening of symptoms requiring new therapy after resolution of anaphylaxis. Significant biphasic reactions were defined as those requiring oxygen, vasopressors, intubation, subcutaneous epinephrine, or unscheduled bronchodilator treatments. Patients were considered to benefit from a 24-hour observation period if they had a significant biphasic reaction within 24 hours of admission. RESULTS: Of 108 anaphylactic episodes, 2 (2%) were fatal, and 1 (1%) was a protracted anaphylactic reaction. Among the remaining 105 children with resolution of anaphylaxis, 6 (6%) [95% confidence (CI): 2, 12] had biphasic reactions, of which 3 (3%) [95% CI:.6, 8] were significant. Of those who had a biphasic reactions, the median time from the onset of symptoms to the initial administration of subcutaneous epinephrine was 190 minutes, versus 48 minutes for those without a biphasic reaction. Patients with or without biphasic reactions did not differ significantly in the incidence of initial epinephrine use, initial steroid use, or serious respiratory or cardiovascular symptoms on initial presentation. Two of 105 (2%) [95% CI:.2, 7] patients clinically benefitted from a 24-hour observation period. CONCLUSIONS: We found an overall incidence of biphasic reactions of 6%, and an incidence of significant biphasic reactions of 3%, among pediatric patients admitted with anaphylaxis. Delayed administration of subcutaneous epinephrine was associated with an increased incidence of biphasic reactions. Approximately 2% of patients with anaphylaxis potentially benefitted from a 24-hour period of observation after symptoms had resolved.

Adolescent↗