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Biomedical subjects

S Lee

Publications and source records attributed to S Lee.

At least 505 records · Page 28Linked to original sources

Experience with the advanced breast biopsy instrumentation system.

OBJECTIVES: To report early experience with the advanced breast biopsy instrumentation (ABBI) system and to compare the results with those of other published studies. DESIGN: A nonrandomized case series. SETTING: An outpatient breast diagnostic centre at a large urban community hospital. PATIENTS: Thirty-four women; 27 had suspicious calcifications, 2 had a nonpalpable mass and 5 had both. INTERVENTION: The ABBI procedure to excise a breast lesion or obtain a representative sample for histologic examination. MAIN OUTCOME MEASURES: Success of the procedure with respect to diagnosis, sample quality, technical problems, margins of tumour free tissue and patient satisfaction. RESULTS: Malignant tissue was diagnosed in 7 women (21%) and atypical ductal hyperplasia in 2 (6%). In all cancers, the obtained samples had malignant cells present at the margins or less than 1 mm away. Technical problems were encountered in 32% of cases. Manual extraction of the specimen was required in 21% of cases. CONCLUSIONS: The preliminary data correlate well with those of other published results. Although it is possible that a small number of cases and a relatively high proportion of technical difficulties may represent a normal learning curve, there is a definite need for improvement of some ABBI components. ABBI does not appear to provide adequate margins of uninvolved tissue in patients with cancer and thus should not be used with curative intent. ABBI provides excellent quality samples for pathological study and good patient satisfaction. There are not yet enough data for meaningful comparison of ABBI with stereotactic core biopsy and excisional biopsy with needle localization.

Adult↗

Left-sided gallbladder associated with anomalous branching of the portal vein detected by sonography.

We experienced a patient with a left-sided gallbladder associated with a right umbilical portion. The etiology of the left-sided gallbladder associated with a right umbilcal portion was in this case not anomalous positioning of the gallbladder, but an embryologic abnormality of the umbilical vein. Ramification of the portal vein should be examined in patients with left-sided gallbladder, especially those scheduled to undergo hepato-biliary surgery, since misunderstanding of anatomy may result in serious problems.

Adult↗

Toward a reverse osmosis membrane system for recycling space mission wastewater.

Essential to extended human exploration and utilization of space is providing a clean supply of potable water as well as water for washing. Recycling of space mission wastewater is necessary for long-term space missions due to the limited capacity of water storage. In this study, initial measurements toward a wastewater reclamation system that provides a clean water supply using reverse osmosis (RO) membranes have been made using stirred cell filtration experiments. Low-pressure reverse osmosis (LPRO) membranes were used to obtain high flux of permeate as well as high rejection. Detergent removal was above 99%, and dissolved salt removal was above 90% in single-pass treatment, while total organic carbon (TOC) removal was nearly 80%. Most problematic is nitrogen rejection, which was 74% at best. Comparison of feed water before and after urea hydrolysis shows that the rejection of nitrogen compounds can be increased to 95% by allowing urea hydrolysis to occur. The removal efficiency for nitrogen compounds was also improved by increasing the shear rate near membrane surface. As a result, the product water in two passes could meet the hygiene water requirements for human space missions, and the product water in three passes could meet potable water regulations with overall recovery of 77%. This study also suggests that dynamic rotating membrane filtration, which can produce a high shear rate, will be useful to increase the system recovery as well as pollutant rejection. Grant numbers: NAG9-1053

Ecological Systems, Closed↗

Isolation of rice allergenic cDNA clones from a rice cDNA library by immunoscreening with a polyclonal antibody specific to 16 kD rice allergenic protein.

Clinical cases of type-1 hypersensitive reaction to rice (Oryza sativa) have been reported in western countries as well as in Japan. Among rice proteins, 14-16 kD globulin proteins encoded by multiple gene family have been identified as major rice allergens. In this study, a rice cDNA library was constructed using lambda UniZap vector and screened with a rat anti-16 kD globulin protein polyclonal antibody in order to isolate Korean rice allergenic cDNA clones. Five independent cDNA clones, termed RAK1-5, were obtained after second rounds of plaque assay and immunoblot analysis. These clones encoded 13-19 kD recombinant proteins upon IPTG induction, which were identified by the polyclonal antibody in immunoblot analysis. DNA sequencing analysis showed that RAK1-4 have 99% sequence homology with RA5b, and RAK5 is closely related with RA14c. This result indicated that RA5b gene is widely distributed in our cDNA library among other possible rice allergenic genes, and more study is needed to isolate heterogeneous or novel rice allergen genes.

Allergens↗

Impaired nucleotide excision repair in UV-irradiated human oral keratinocytes immortalized with type 16 human papillomavirus genome.

We previously reported that 'high risk' human papillomaviruses (HPV) induce genetic instability in human oral keratinocytes. To understand the mechanisms of HPV-induced genetic instability, we determined the nucleotide excision repair (NER) capacity of normal (NHOK) and human papillomavirus type-16 immortalized oral keratinocytes (HOK-16B) by strand-specific removal of UV-induced cyclobutane pyrimidine dimers (CPDs) from a 16 Kb fragment of the p53 gene. In NHOK the NER activity was initiated in both DNA strands immediately, although the process in the non-transcribed strand was notably slower than that of the transcribed strand. In HOK-16B cells the initiation of CPDs removal was delayed for at least 8 h in both DNA strands, and the process was significantly slower than that in NHOK. UV-irradiation enhanced the p53 protein level more than 30-fold in NHOK, but it did not significantly alter the protein level in the HOK-16B cells. UV-irradiation also increased the p21WAF1/CIP1 protein level only in NHOK. These data indicate that 'high risk' HPV induces genetic instability by impairing NER capacity of cells. Impaired NER activity of HOK-16B cells may be implicated with their inability to enhance active p53 when challenged by genotoxic stress.

Cell Line, Transformed↗

Lack of differences in nef alleles among HIV-infected asymptomatic long-term survivors and those who progressed to disease.

Sequences of the human immunodeficiency virus (HIV) nef gene in virus isolates from 12 long-term survivors (LTSs) and 7 progressors were compared to determine if any association existed between the sequences and the corresponding clinical status. The sequences of at least five clones were determined for each subject. Conceptual translations of the open reading frames (ORFs) were examined with respect to a consensus with a prototypic nef sequence (HIV-1SF2) and for conservation of functionally described motifs. Premature stops were observed at equivalent, yet low, frequencies among the different clinical groups: 2 of 60 (3.33%) and 1 of 45 (2.22%) respectively. No remarkable differences in protein motifs implicated in several activities ascribed to Nef were noted. An association between nef sequence characteristics and the clinical state was not found.

Alleles↗

Inhibition of initiation of simian virus 40 DNA replication in vitro by the ursodeoxycholic acid and its derivatives.

In this study, the effects of the ursodeoxycholic acid (UDCA), and its derivatives, on DNA replication were examined using simian virus (SV40) DNA replication in vitro. We found that UDCA and its derivatives inhibited SV40 DNA replication, and predominantly inhibited the initiation stage of DNA replication. UDCA and its derivatives inhibited the DNA cleavage by topoisomerase I (topo I). Among them, HS-1183 significantly reduced the activity of topo I. UDCA, at 100 microM, significantly reduced polymerase alpha-primase (pol alpha-primase) activity, but HS-1030 and HS-1183 showed a weak inhibitory effect. The ssDNA binding activity of replication protein A (RPA) was little affected by UDCA and HS-1030, but was weakly inhibited by HS-1183. Based on their properties, we suggest that UDCA and its derivatives might inhibit some molecules that is required to establish replication forks during the initiation reaction and their cytotoxicity might be related to the inhibitory effect they have on this fundamental cellular process.

Antineoplastic Agents↗

Intracellular assembly of Kell and XK blood group proteins.

Kell, a 93 kDa type II membrane glycoprotein, and XK, a 444 amino acid multi-pass membrane protein, are blood group proteins that exist as a disulfide-bonded complex on human red cells. The mechanism of Kell/XK assembly was studied in transfected COS cells co-expressing Kell and XK proteins. Time course studies combined with endonuclease-H treatment and cell fractionation showed that Kell and XK are assembled in the endoplasmic reticulum. At later times the Kell component of the complex was not cleaved by endonuclease-H, indicating N-linked oligosaccharide processing and transport of the complex to a Golgi and/or a post-Golgi cell fraction. Surface-labeling of transfected COS cells, expressing both Kell and XK, demonstrated that the Kell/XK complex travels to the plasma membrane. XK expressed in the absence of Kell was also transported to the cell surface indicating that linkage of Kell and XK is not obligatory for cell surface expression.

Amino Acid Transport Systems, Neutral↗

Solution structure of neuromedin B by (1)H nuclear magnetic resonance spectroscopy.

The solution structure of neuromedin B (NMB) was investigated using two-dimensional nuclear magnetic resonance (NMR) spectroscopy in membrane-mimicking environments. NMB adopts a relaxed helical conformation from Trp(4) to Met(10) in 50% aqueous 2,2, 2-trifluoroethanol (TFE) solution and in 150 mM SDS micelles. Sidechain atoms of the three residues, Trp(4), His(8) and Phe(9) orient toward the same direction and these residues might play a key role on interacting with hydrophobic acyl chains of the phospholipids in the membrane. NOESY experiments performed on NMB in non-deuterated SDS micelle show that aromatic ring protons of Trp(4) and Phe(9) residues are in close contact with methylene protons of SDS micelles. In addition, proton longitudinal relaxation data proved that the interactions between NMB with SDS micelle are characterized as extrinsic interaction. Trp(4) and Phe(9) seem to be important in interaction with receptor and this agrees with the previous studies of structure-activity relationship (Howell, D.C. et al. (1996) Int. J. Pept. Protein Res. 48, 522-531). These conformational features might be helpful in understanding the molecular mechanism of the function of NMB and developing the efficient drugs.

Histidine↗

Extraosseous extension of vertebral hemangioma, a rare cause of spinal cord compression.

STUDY DESIGN: Case report. OBJECTIVE: To illustrate a rare cause of thoracic spinal cord compression, its diagnosis, and its management. SUMMARY OF BACKGROUND DATA: Asymptomatic vertebral hemangiomas are common, but extraosseous extension causing spinal cord compression with neurologic symptoms is rare, and few cases appear in the English-language literature. METHOD: A previously asymptomatic 63-year-old man sought medical attention for acute back pain and thoracic myelopathy of 6 week's duration. Magnetic resonance imaging confirmed the presence of a mass in the T10 vertebral body with paravertebral and intracanalicular extension contributing to cord compression. Decompression and reconstructive surgery were performed and radiotherapy administered after surgery. Preoperative angiography was not performed because of the patient's rapidly progressive neurologic deterioration and the consideration that the differential diagnosis of vertebral hemangioma was less likely. RESULTS: The diagnosis of benign capillary hemangioma was made histologically. Neurologic recovery was complete except for minor residual sensory changes in the legs. At follow-up 10 months after surgery the patient had returned to his usual active life and motor mower repairing business. CONCLUSION: Extraosseous extension of vertebral hemangiomas is a rare cause of thoracic spinal cord compression. As such, the available data are derived from reports based on series involving only a small number of cases, rather than on results of randomized controlled trials. Those causing progressive neurologic symptoms should be surgically decompressed, with the specific procedure determined by the extent and site of the lesion. Preoperative angiography is recommended, but embolization is not always necessary or even possible. Postoperative radiotherapy is recommended when tumor removal is subtotal.

Angiography↗

Identification of residues in the monocyte chemotactic protein-1 that contact the MCP-1 receptor, CCR2.

The CC chemokine, MCP-1, has been identified as a major chemoattractant for T cells and monocytes, and plays a significant role in the pathology of inflammatory diseases. To identify the regions of MCP-1 that contact its receptor, CCR2, we substituted all surface-exposed residues with alanine. Some residues were also mutated to other amino acids to identify the importance of charge, hydrophobicity, or aromaticity at specific positions. The binding affinity of each mutant for CCR2 was assayed with THP-1 and CCR2-transfected CHL cells. The majority of point mutations had no effect. Residues at the N-terminus of the protein, known to be crucial for signaling, contribute less than a factor of 10 to the binding affinity. However, two clusters of primarily basic residues (R24, K35, K38, K49, and Y13), separated by a 35 A hydrophobic groove, reduced the level of binding by 15-100-fold. A peptide fragment encompassing residues 13-35 recapitulated some of the mutational data derived from the intact protein. It exhibited modest binding as a linear peptide and dramatically improved affinity when the region which adopts a single turn of a 3(10)-helix in the protein, which includes R24, was constrained by a disulfide bond. Additional constraints at the ends of the peptide, corresponding to the disulfide between the first and third cysteines in MCP-1, yielded further improvements in affinity. Together, these data suggest a model in which a large surface area of MCP-1 contacts the receptor, and the accumulation of a number of weak interactions results in the 35 pM affinity observed for the wild-type (WT) protein. The receptor binding site of MCP-1 also is significantly different from the binding sites of RANTES and IL-8, providing insight into the issue of receptor specificity. It was previously shown that the N-terminus of CCR2 is critical for binding MCP-1 [Monteclaro, F. S., and Charo, I. F. (1996) J. Biol. Chem. 271, 19084-92; Monteclaro, F. S., and Charo, I. F. (1997) J. Biol. Chem. 272, 23186-90]. Point mutations of six acidic residues in this region of the receptor were made to test their role in ligand binding. This identified D25 and D27 of the DYDY motif as being important. On the basis of our data, we propose a model in which the receptor N-terminus lies along the hydrophobic groove in an extended fashion, placing the DYDY motif near the basic cluster involving R24 and K49 of MCP-1. This in turn orients the signaling residues (Y13 and the N-terminus) for productive interaction with the receptor.

Amino Acid Sequence↗

Distribution fate and mechanism of immune modulation following mucosal delivery of plasmid DNA encoding IL-10.

DNA vaccination has been widely studied in several models of vaccination and in the treatment of inflammatory diseases, even though the mechanism involved is still unclear. This report demonstrates that mucosal administration of plasmid DNA leads to rapid and widespread distribution around the body. Dissemination likely occurred via the bloodstream because plasmid DNA was present in blood plasma. The plasmid DNA was also detectable in several tissues including draining lymph nodes, spleen, liver, bone marrow, and even the dermis of ear pinnae. Except for the site of administration, plasmid DNA was no longer detectable in tissues after 3 wk postadministration. RNA and protein expression was also found in the tissues and bloodstream. Animals previously primed by HSV infection and subsequently given IL-10 DNA via the nasal mucosa, showed diminished Ag-induced delayed type hypersensitivity reactions for up to 5 wk posttreatment. The mechanism of modulation involved diminished the Ag-specific proliferation and production of Th1 cytokines. The Ag-specific silencing effects persisted beyond the duration of detectable plasmid encoded protein and was maintained upon adoptive transfer of T cells into a plasmid-free environment. The silenced T cells were not a source of IL-10, and their anergic state was reversible by exposure to Ag in the presence of exogenous IL-2.

Adjuvants, Immunologic↗

Nitric oxide stimulates ACTH secretion and the transcription of the genes encoding for NGFI-B, corticotropin-releasing factor, corticotropin-releasing factor receptor type 1, and vasopressin in the hypothalamus of the intact rat.

We investigated the effect of the intracerebroventricular injection of the nitric oxide (NO) donor 3-morpholino-sydnonimine (SIN-1) on the release of adrenocorticotropin hormone (ACTH) and the neuronal response of hypothalamic neurons responsible for this release. Rats that were administered SIN-1 showed significant elevations in plasma ACTH levels, a response that was virtually abolished by antibodies against corticotropin-releasing factor (CRF) and significantly blunted by vasopressin (VP) antiserum. SIN-1 also upregulated heteronuclear (hn) transcripts for CRF and VP and messenger RNA (mRNA) levels for the immediate early gene NGFI-B and for CRF receptor type 1 (CRF-R(1)) in the parvocellular portion of the paraventricular nucleus (PVN) of the hypothalamus. Blockade of prostaglandin synthesis with ibuprofen did not alter the ACTH or the PVN response to SIN-1. The central nucleus of the amygdala and the supraoptic nucleus, regions that are involved in autonomic adjustments to altered cardiovascular activity, also responded to SIN-1 with elevated NGFI-B mRNA levels. However, the only change in mean arterial blood pressure caused by this NO donor was a transient and modest increase. To our knowledge, this is the first demonstration that in the intact rat NO stimulates the activity of PVN neurons that control the hypothalamic-pituitary-adrenal axis. It must be noted, however, that our results do not allow us to determine whether this effect was direct or mediated through PVN afferents. This study should help resolve the controversy generated by the use of isolated brain tissues to investigate the net effect of NO on hypothalamic peptide production.

Adrenocorticotropic Hormone↗

Neonatal footshock stress alters adult behavior and hippocampal corticosteroid receptors.

To determine the effects of stress early in life on adult behavior and hippocampal corticosteroid receptors, rats were exposed to footshocks (0.8 mA, 60 times/day, randomly apart) on postnatal days 14, 17 and 20. When they reached 6 months of age, neurobehavioral alterations were measured. The footshock-experienced rats learned more rapidly in the autoshaped learning test than similarly handled controls. They also stabilized more quickly after exposure to a novel environment than the handled controls, but only at the same rate as animals which had not been handled except for weighing. The density of [3H]dexamethasone binding sites increased and that of [3H]corticosterone binding sites decreased in the hippocampi of these rats. These results indicate that early life stress results in altered behavior and hippocampal corticosteroid receptors at adulthood, and suggest that the mineralocorticoid and the glucocorticoid receptors are differentially regulated by early life stress.

Animals↗