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Biomedical subjects

J Lu

Publications and source records attributed to J Lu.

At least 487 records · Page 27Linked to original sources

Liver injury model in mice induced by a cellular immunologic mechanism--delayed-type hypersensitivity-induced liver injury to picryl chloride and phenotype of effector cell.

Liver injury was induced in BALB/c mice by local delayed-type hypersensitivity (DTH) to picryl chloride (PC1). Distinct changes of biochemical parameters were observed including the elevation of serum alanine and aspartate aminotransferases, increase of liver lipid peroxides, as well as decrease of serum alkaline phosphatase. Damage was confirmed by histopathological findings such as hepatocellular necrosis, granulocyte infiltration, and fatty degeneration. The liver injury was passively transferred into naive syngeneic mice by infusing spleen cells from immune mice. The capacity of the splenocytes to induce liver injury in recipient mice was almost completely abolished by pretreatment of the cells with anti-Thy 1.2 or anti-CD4, but not anti-CD8 antibody. These findings suggest that the production of liver injury by a local DTH mechanism is possible and the subpopulation of T cells, Thy-1.2+, L3T4+, and Lyt-2- cells, is at least one of the effector cells that mediate the injury.

Animals↗

The effect of the pediatric clerkship on medical student attitudes toward pediatrics at 11 medical schools.

OBJECTIVES: To investigate how the pediatric clerkship affected student attitudes toward pediatrics, and to determine if correlations existed between changes in attitudes toward pediatrics and in ratings of certain aspects of the clerkship with an increased interest in a pediatric career. METHODS: A one-page survey measuring interest in a career in pediatrics and agreement or disagreement with seven statements about pediatrics was administered at the beginning and end of the pediatric clerkship at 11 medical schools for the 1992-1993 academic year. RESULTS: The proportion of students with a strong interest in a pediatric career increased from 6.7% before the clerkship to 15.2% after the clerkship (for women, 11% to 22%; for men, 4% to 11%). Attitudes toward pediatrics were more favorable at the end vs the beginning of the clerkship. The change that correlated best with change in interest in a pediatrics career was agreement that children are enjoyable to work with. Of the eight aspects of the clerkship rated, the patients worked with on the ward received the most positive mean score. The item that correlated best with increased career interest was a positive feeling toward the ward residents. CONCLUSION: The recent trend for women to have a greater interest in careers in pediatrics than men is continuing. Finding ways to make students more comfortable when they interact with children and improving the teaching skills of residents could improve recruitment of medical students into pediatrics.

Career Choice↗

Contribution of the tyrosines to the structure and function of the human U1A N-terminal RNA binding domain.

RNA binding domains (RBDs) are members of a large family of proteins that share minimal sequence conservation but adopt an alpha beta sandwich global fold. Defining the contributions of specific amino acids to RBD structure and RNA binding is critical to understanding the functions of these proteins. In these experiments with the human U1A N-terminal RNA binding domain (RBD1), the contributions from each of its four tyrosines to protein structure, stability, and RNA binding were measured. Each tyrosine was substituted with phenylalanine and one other selected residue, and the resulting proteins were characterized by chemical denaturation to measure their unfolding free energy, by binding free energies to the wild-type RNA hairpin, and by 19F NMR to probe for structural changes. Features of the protein identified in these experiments include a possible tyrosine/lysine contact in an alpha-helix, which may be an example of an energetically favorable aromatic/amino side chain interaction. One long loop of the protein, which shows unusual 15N backbone and tyrosine side-chain dynamics, is implicated in protein:protein association. The diverse interactions of the four tyrosine residues in the organization of RBD1 illustrate how each member of this family of proteins will have unique molecular details that contribute to function.

Amino Acid Sequence↗

IA-2 and IA-2 beta are major autoantigens in IDDM and the precursors of the 40 kDa and 37 kDa tryptic fragments.

By subtraction strategy and polymerase chain reaction amplification, two novel cDNAs, designated IA-2 and IA-2 beta, were cloned, sequenced and expressed. Both are transmembrane proteins belonging to the protein tyrosine phosphatase family and are expressed in pancreatic islets. Serological studies revealed that a high percentage of patients with IDDM have autoantibodies to IA-2/IA-2 beta and that the presence of these autoantibodies in otherwise normal individuals is highly predictive in identifying those at risk of ultimately developing clinical diabetes. Moreover, many patients who are ICA positive, but who do not have Abs to GAD65, have Abs to IA-2/IA-2 beta. Enzymatic cleavage of IA-2/IA-2 beta and serological analysis showed that IA-2 is the precursor of the 40 kDa tryptic fragment and IA-2 beta is the precursor of the 37 kDa tryptic fragment, both previously shown to be autoantigens. It is concluded that IA-2/IA-2 beta are major autoantigens in IDDM and together with GAD65 are responsible for much of the reactivity of ICA with pancreatic islets. Tests for the detection of autoantibodies to recombinant IA-2/IA-2 beta and recombinant GAD65 are likely to replace the ICA immunofluorescence test for population screening.

Amino Acid Sequence↗

A modified dorsal approach to the wrist for arthrodesis of the non-rheumatoid wrist. An anatomical study.

Fourteen cadaveric wrists were dissected to investigate a modified dorsal approach that involved osteotomy of the small and dorsal tubercles without opening the third compartment. This approach could be safely made with good exposure of the dorsum of the wrist. The mean normal angle formed by the extensor pollicis longus tendon at the level of the dorsal tubercle was 144 degrees. An approach that involves division of the third compartment may lead to effective lengthening of the extensor pollicis longus musculotendinous unit by 8 to 17 mm with corresponding decrease in the tension generated by its contraction. The modified approach permits restoration of the normal alignment of the extensor pollicis longus tendon, and may be useful for performing arthrodesis of the non-rheumatoid wrist in young manual workers.

Arthritis, Rheumatoid↗

The agreement between ventricular volumes and ejection fraction by transesophageal echocardiography or a combined radionuclear and thermodilution technique in patients after coronary artery surgery.

OBJECTIVES: To study the reproducibility of and agreement between perioperative transesophageal echocardiographic (TEE) and radionuclide (RN) assessment of ventricular volumes and ejection fraction (EF). DESIGN: A prospective, blinded comparison of two methods of measurement. SETTING: A surgical intensive care unit in a university hospital. PARTICIPANTS: Patients after coronary artery bypass surgery. INTERVENTIONS: Left ventricular volumes and ejection fraction were simultaneously measured by radionuclide ventriculography and transesophageal echocardiography. Two sets of measurements were made in rapid succession with a third set after an interval. RESULTS: Reproducibilities of EF and ventricular volumes by TEE and RN were similar. At each set of measurements, the bias for radionuclide EF and TEE Simpson's rule EF, 0.03 +/- 0.05, 0 +/- 0.06, -0.01 +/- 0.07, respectively, for radionuclide EF and TEE area length EF 0.01 +/- 0.05, -0.01 +/- 0.05, -0.03 +/- 0.08, respectively, were significantly less than for radionuclide EF and TEE FAC 0.07 +/- 0.05, 0.05 +/- 0.05, 0.03 +/- 0.09. Poor agreement was observed between RNTD-EDV, and both of the TEE EDV measurements. CONCLUSION: EF measured by TEE area length and Simpson's rule method are as reproducible as TEE FAC and are more accurate estimates of RN EF. Poor agreement between methods of measuring end-diastolic volume was observed.

Aged↗

Biosynthesis of human ficolin, an Escherichia coli-binding protein, by monocytes: comparison with the synthesis of two macrophage-specific proteins, C1q and the mannose receptor.

Ficolin is characterized by the presence of both collagen-like and fibrinogen-like sequences, and potentially has a similar overall structure as the complement protein C1q and the collectins. Previous studies have reported the presence of human ficolin mRNA predominantly in peripheral blood leucocytes. In the present study, the cellular origin of human ficolin was investigated in further detail. Preliminary studies using reverse transcriptase-polymerase chain reaction (RT-PCR) showed that ficolin mRNA was synthesized by U937 cells, a human monocyte cell line. This finding suggested that blood monocytes also normally synthesize human ficolin. Peripheral blood monocytes from adult human donors were harvested at serial time-points (0-20 hr) after adhesion to tissue culture plates, and total RNA was isolated and assayed for ficolin mRNA by RT-PCR. Ficolin mRNA was highly expressed in monocytes throughout the first 20 hr of adhesion. In contrast, C1q and mannose receptor mRNA were not detectable during the first 8 hr of adhesion, but were highly expressed by 20 hr. Cells were harvested at longer time intervals (1, 2, 4, 6 and 8 days) to determine whether ficolin expression was temporally regulated at later stages of monocyte differentiation. Ficolin mRNA levels decreased sharply from day 1 to day 6. In contrast, the levels of both C1q and mannose receptor mRNA showed no changing trend. These results are consistent with the absence of ficolin expression in many macrophage-rich tissues previously reported. The origin of ficolin from monocytes, together with its structural similarity to C1q and the collectins, raises the possibility that ficolin may be another plasma protein capable of binding to surface structures of micro-organisms. Escherichia coli was therefore incubated with human serum, and bound proteins, after elution with sugars, were analysed by Western blotting using an antiserum raised against a synthetic ficolin peptide. The antiserum identified a polypeptide of approximately 42000 MW, which is similar in size to that of ficolin as predicted from its cDNA-derived sequence.

Blotting, Western↗

Characterization and chromosomal localization of a new protein disulfide isomerase, PDIp, highly expressed in human pancreas.

Protein disulfide isomerase (PDI) catalyzes protein folding and thiol-disulfide interchange reactions. The enzyme is localized in the lumen of endoplasmic reticulum (ER) and is abundant in secretory cells of various tissues. In this study we describe the isolation and characterization from human pancreas of a new protein, PDIp, that is structurally and functionally related to PDIs. PDIp cDNA is 1,659 bp in length and predicts a protein with an open reading frame of 511 amino acids. PDIp amino acid sequence shows 46% identity and 66% similarity to that of human PDI. PDIp possesses two thioredoxin-like active sites (WCGHCQ and WCTHCK) and an endoplasmic reticulum retention signal sequence, KEEL, at the carboxyl terminus. Northern analysis of normal human tissues and various human tumor cell lines revealed PDIp mRNA (2.0 kb) expression only in the normal pancreas. Recombinant PDIp protein catalyzed reductive cleavage of insulin and renaturation of reduced RNaseA. Somatic cell genetics and fluorescence in situ hybridization localized the PDIp gene to the short arm of human chromosome 16. It is concluded that PDIp is a new member of the PDI family and is highly expressed in human pancreas.

Amino Acid Sequence↗

Effect on an aqueous extract of selenium-enriched garlic on in vitro markers and in vivo efficacy in cancer prevention.

Previous work has shown that the efficacy of cancer prevention by selenium-enriched garlic (Se-garlic) is primarily dependent on the action of selenium. An aqueous extract containing 43 micro Se/ml was prepared from lyophilized Se-garlic powder by the Soxhlet method. The activity of this Se-garlic extract was evaluated in a transformed mammary epithelial cell culture model for its effect on cell morphology, cell growth, cell cycle progression and the induction of single and double stranded breaks in DNA. Comparisons were also made with a similarly prepared extract from regular garlic, Se-methylselenocysteine (a major water-soluble seleno-amino acid identified in Se-garlic) and selenite (used for fertilizing Se-garlic). In contrast to the regular garlic extract which produced little or no modulation of the above parameters, treatment with the Se-garlic extract resulted in growth inhibition, GI phase cell cycle arrest and apoptotic DNA double strand breaks in the absence of DNA single strand breaks. This pattern of cellular responses was duplicated with exposure to Se-methylselenocysteine. Selenite, on the other hand, induced cell cycle blockage in the S/G2-M phase, and a marked increase in DNA single strand breaks (a measure of genotoxicity) in addition to growth suppression. The chemopreventive efficacy of the two garlic extracts was also investigated in the rat methylnitrosourea mammary tumor model. Both extracts were supplemented in the diet for 1 month immediately following carcinogen administration. Significant cancer protection was observed with treatment by the Se-garlic extract (at 3 p.p.m. Se in the diet), while little benefit was noted with treatment by the regular garlic extract. Based on the above in vitro and in vivo findings, it is hypothesized that the Se-garlic extract, in part via the action of Se-methylselenocysteine, is able to inhibit tumorigenesis by suppressing the proliferation and reducing the survival of the early transformed cells. Furthermore, the data also support the concept that the modulation of certain in vitro markers may be of value in predicting the effectiveness of novel forms of selenium for cancer prevention.

Animals↗

Thoracic spine fractures in patients older than 50 years.

The charts and radiographs of 70 patients older than 50 years of age with thoracic vertebral body collapse were reviewed retrospectively. Fifteen patients had traumatic fractures and 34 had osteoporotic collapse of thoracic vertebrae. Metastasis was the underlying disease process in 18 patients and multiple myeloma in the remaining 3 patients. Thirteen patients had fractures involving the upper half of the dorsal spine, of which 8 (61.5%) were metastatic, 4 (30.8%) osteoporotic, and 1 (7.7%) traumatic. All patients with osteoporotic fractures of the upper dorsal spine also had 1 or more fractures of the lower dorsal or lumbar spine. There were 11 metastatic, 80 osteoporotic, 14 traumatic, and 3 fractures secondary to multiple myeloma involving the lower dorsal spine. There were no infections or primary bone tumors. The difference in the frequency of metastatic fractures against other etiologies involving the upper versus the lower thoracic spine was highly statistically significant.

Aged↗

A gene-rich cluster between the CD4 and triosephosphate isomerase genes at human chromosome 12p13.

The genomic sequence of the human CD4 gene and its neighboring region, located at chromosome 12p13, was generated using the large-scale shotgun sequencing strategy. A total of 117 kb of genomic sequence and approximately 11 kb of cDNA sequence were obtained. Six genes, including CD4, triosephosphate isomerase, B3 subunit of G proteins (GNB3), and ubiquitin isopeptidase T (ISOT), with known functions, and two new genes with unknown functions were identified. Using a battery of strategies, the exon/intron boundaries, splice variants, and tissue expression patterns of the genes were determined. Various computer software was utilized for analyses of the DNA and amino acid sequences. The results of the analyses and sequence-based strategies for gene identification are discussed.

Alternative Splicing↗

Computer evaluation of second tarsometatarsal joint dislocation.

Using computer-assisted techniques, this study analyzes the mean contact area of the articular surface of the second tarsometatarsal joint. The articular contact area decreased proportionate to the displacement in both males and females, but it was consistently greater in males than in females for all simulated displacements. The reduction in the contact area was the highest with dorsolateral displacement compared with the lateral and dorsal displacements. Dorsolateral displacement of the second metatarsal of 3 mm led to 38.6% reduction in the contact area, compared with 33.1% and 20.2% reduction with lateral and dorsal displacements, respectively. This study shows that even minor degrees of displacement not apparent on plain radiographs lead to significant decrease in the contact area of the second tarsometatarsal joint. Careful evaluation of second tarsometatarsal injuries with computed tomography is recommended to detect minor degrees of displacement.

Computer Simulation↗

Method for 96-well M13 DNA template preparations for large-scale sequencing.

Efficient preparation of DNA templates is an important step in large-scale DNA sequencing. The ensure high-quality sequence data, we have prepared M13 phage DNA templates using a glass fiber-filtration method. We present the adaptation of this protocol to a 96-well format using commercially available filter plates. Two variations are described: one using polyethylene glycol precipitation and a second where the phage particles are disrupted before filtration, thus eliminating the need for precipitation. Using either of these protocols, 96 templates can be prepared in less than 2 h. Sufficient DNA for 1-2 dye primer sequencing reactions is routinely obtained from 1 mL of culture, and the resulting sequence data are of high quality.

Bacteriophage M13↗

Vulnerability of vertebral artery in anterolateral decompression for cervical spondylosis.

This study aimed to provide anatomic data for the location of the vertebral artery and offer an optimal approach for lateral cervical decompression that minimizes the risk of injury to the vertebral artery. Anatomically, there has been little study documenting the safe zone to prevent vertebral artery injury during the resection of the uncinate process or uncovertebral joint during the lateral decompression of the nerve root. The transverse foramen and its related parameters were measured on dry cervical spines from C3 to C7. The cadaveric cervical spines were dissected to determine a method for resection of the uncovertebral joint with decreased risk of vertebral artery laceration. The anteroposterior diameters of the transverse foramina gradually decreased from C6 to C3. The transverse diameters of the transverse foramina were smaller at C5. The interforaminal distance, width of the vertebrae, interuncinate distance, and the distance from the lateral tip of the uncinate process to the medial border of the transverse foramen became smaller in more cephalad vertebrae. After subtotal vertebrectomy and opening of the anterior walls of the transverse foramina, the resection of the uncovertebral joint and lateral decompression became easier and safer. Anatomic measurements obtained in this study indicate the vertebral artery to be at risk during decompression of the more cephalad vertebrae. The lateral decompression can be completed under direct vision with smaller rongeurs and curettes, rather than with high speed burr after deroofing the anterior walls of transverse foramina and retracting the vertebral artery laterally.

Adult↗

An immunohistochemical study of the intraventricular macrophages in induced hydrocephalus in prenatal rats following a maternal injection of 6-aminonicotinamide.

Hydrocephalus was induced experimentally in prenatal rats following an injection of 6-aminonicotinamide (6-AN) into pregnant rats. The most remarkable change of the dilated lateral ventricles was in a marked increase in the number of intraventricular macrophages, some of which were laden with ingested erythrocytes. The immunoreactivity of the intraventricular macrophages was noticeably enhanced with the monoclonal antibodies OX-42 and OX-18 which marked the complement type 3 receptors (CR3) and major histocompatibility complex (MHC) class I antigen, respectively. Many immunoreactive cells with similar external morphology were observed to penetrate the ependymal lining at the roof of the ventricles. This, coupled with the concomitant depletion of the conglomeration of amoeboid microglia in the supraventricular corpus callosum, suggests that the upsurge of immunoreactive intraventricular macrophages in hydrocephalus was partly due to the influx of amoeboid microglia probably in response to the damage of the ventricular walls and possible alteration in the contents of the cerebrospinal fluid. The significance of the upregulation of complement type 3 receptors and major histocompatibility complex class I antigens on epiplexus cells in hydrocephalic rats remains to be explored, although our results suggest that the surface antigens may be involved in increased phagocytosis and/or a possible immune response.

6-Aminonicotinamide↗