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Biomedical subjects

P Tan

Publications and source records attributed to P Tan.

At least 37 records · Page 2Linked to original sources
PubMed2000

Biocompatibility of electroactive polymers in tissues.

The biocompatibility of ethylene-vinyl acetate copolymer (EVAc), polyethylene (PE), and polyaniline (PANi) films in the emeraldine (EM), nigraniline (NA) and leucoemeraldine (LM) intrinsic oxidation states were assessed through subcutaneous implantation into male Sprague-Dawley rats beneath the dorsal skin, for a period ranging from 19 to 90 weeks. Histological examination, interstitial pressure measurement, and X-ray photoelectron spectroscopy (XPS) were employed to determine the biocompatibility of the polymers. The polymers did not provoke inflammatory responses in the subcutaneous tissues over the entire implantation period. Characteristics features associated with tissue-implant incompatibility were not evident near the implantation. Interstitial pressure was measured to evaluate the development of tissue. Low interstitial pressure readings on the region of implantation confirmed the biocompatibility of these polymer types. The surface composition of the electroactive aniline polymers before and after the implantation was characterized by XPS.

Aniline Compounds↗
PubMed2000

"Fluorescent timer": protein that changes color with time.

We generated a mutant of the red fluorescent protein drFP583. The mutant (E5) changes its fluorescence from green to red over time. The rate of color conversion is independent of protein concentration and therefore can be used to trace time-dependent expression. We used in vivo labeling with E5 to measure expression from the heat shock-dependent promoter in Caenorhabditis elegans and from the Otx-2 promoter in developing Xenopus embryos. Thus, E5 is a "fluorescent timer" that can be used to monitor both activation and down-regulation of target promoters on the whole-organism scale.

Animals↗
PubMed2000

The conserved RING-H2 finger of ROC1 is required for ubiquitin ligation.

ROC1 is a common component of a large family of ubiquitin E3 ligases that regulate cell cycle progression and signal transduction pathways. Here we present evidence suggesting that a conserved RING-H2 structure within ROC1 is critical for its ubiquitin ligation function. Mercury-containing sulfhydryl modification agents (rho-hydroxymercuribenzoate and mercuric chloride) irreversibly inhibit the ROC1-CUL1 ubiquitin ligase activity without disrupting the complex. Consistent with this, these reagents also eliminate the ability of the Skp1-CUL1-HOS-ROC1 E3 ligase complex to support the ubiquitination of IkappaBalpha. Site-directed mutagenesis analysis identifies RING-H2 finger residues Cys(42), Cys(45), Cys(75), His(77), His(80), Cys(83), Cys(94), and Asp(97) as being essential for the ROC1-dependent ubiquitin ligase activity. Furthermore, C42S/C45S and H80A mutations reduce the ability of ROC1 to interact with CUL1 in transfected cells and diminish the capacity of ROC1-CUL1 to form a stable complex with Cdc34 in vitro. However, C75S, H77A, C94S, and D97A substitutions have no detectable effect on ROC1 binding activities. Thus, the ROC1 RING-H2 finger may possess multiple biochemical properties that include stabilizing an interaction with CUL1 and recruiting Cdc34. A possible role of the RING finger in facilitating the Ub transfer reaction is discussed.

Amino Acid Sequence↗
PubMed2000

Molecular engineering of proteins and polymers for targeting and intracellular delivery of therapeutics.

There are many protein and DNA based therapeutics under development in the biotechnology and pharmaceutical industries. Key delivery challenges remain before many of these biomolecular therapeutics reach the clinic. Two important barriers are the effective targeting of drugs to specific tissues and cells and the subsequent intracellular delivery to appropriate cellular compartments. In this review, we summarize protein engineering work aimed at improving the stability and refolding efficiency of antibody fragments used in targeting, and at constructing new streptavidin variants which may offer improved performance in pre-targeting delivery strategies. In addition, we review recent work with pH-responsive polymers that mimic the membrane disruptive properties of viruses and toxins. These polymers could serve as alternatives to fusogenic peptides in gene therapy formulations and to enhance the intracellular delivery of protein therapeutics that function in the cytoplasm.

Antibodies↗
PubMed2000

Impaired immune responses and altered peptide repertoire in tapasin-deficient mice.

Tapasin is a component of the major histocompatibility complex (MHC) class I antigen-loading complex. Here we show that mice with a disrupted tapasin gene display reduced MHC class I expression. Cytotoxic T cell (CTL) responses to viruses are impaired, and dendritic cells of tapasin-deficient mice do not cross-present protein antigen via the MHC class I pathway, indicating a defect in antigen processing. Natural killer (NK) cells from tapasin-deficient mice have an altered repertoire and are self-tolerant. In addition, the repertoire of class I-bound peptides is altered towards less stably binding ones. Thus tapasin plays a role in CTL and NK immune responses and in optimal peptide selection.

Animals↗
PubMed2000

What influences medical students' choice of surgical careers.

BACKGROUND: Recent emphasis in medical education has been to encourage students to pursue primary care careers. This could have a negative impact on applications to surgical residencies. METHODS: To determine what factors are most influential for a student to pursue a surgical career in spite of this environment, third- and fourth-year medical students were surveyed with a 40-item questionnaire. RESULTS: The response rate was 37% (76/205). Those students considering a career in surgery were more likely than their counterparts to be motivated by role models (P <.006), career opportunities (P <.006), and academic opportunities (P <.013) in surgery. They were less likely than their counterparts to be discouraged from surgery on the basis of lifestyle (P <.001), time commitment (P <.001), call schedules (P <.001), or residency length (P <.028). No differences regarding financial rewards, research opportunities, or intellectual challenges were seen between the groups. Neither race nor sex had a significant role in the selection of surgery as a career. CONCLUSIONS: The data suggests that students are more likely to be influenced to pursue surgical careers by offering early exposure to positive role models and career and academic opportunities in surgery. Knowledge of these influences on student career choices should help surgical educators attract and maintain student interest in surgical careers.

Career Choice↗
PubMed2000

Chemical modification of a variant of human MIP-1alpha; implications for dimer structure.

A sequence variant of human MIP-1alpha, in which Asp26 has been replaced by Al alpha, has been chemically modified by the addition of 13C-labeled methyl groups at each of the lysine residues and the N-terminus. The sites of methylation have been verified by a combination of MALDI-TOF mass spectrometric experiments and tryptic digestion followed by N-terminal mapping. The effect of the modification on the structure and activity of the protein have been determined by analytical ultra-centrifugation, 13C NMR spectroscopy and receptor binding studies. The results of these experiments suggest that huMIP-alpha D26A (BB10010), when present as a dimer, adopts a globular structure, like MCP-3, rather than the elongated or cylindrical structure determined for dimers of huMIP-1beta and RANTES.

Alanine↗
PubMed2000

The SCF(HOS/beta-TRCP)-ROC1 E3 ubiquitin ligase utilizes two distinct domains within CUL1 for substrate targeting and ubiquitin ligation.

We describe a purified ubiquitination system capable of rapidly catalyzing the covalent linkage of polyubiquitin chains onto a model substrate, phosphorylated IkappaBalpha. The initial ubiquitin transfer and subsequent polymerization steps of this reaction require the coordinated action of Cdc34 and the SCF(HOS/beta-TRCP)-ROC1 E3 ligase complex, comprised of four subunits (Skp1, cullin 1 [CUL1], HOS/beta-TRCP, and ROC1). Deletion analysis reveals that the N terminus of CUL1 is both necessary and sufficient for binding Skp1 but is devoid of ROC1-binding activity and, hence, is inactive in catalyzing ubiquitin ligation. Consistent with this, introduction of the N-terminal CUL1 polypeptide into cells blocks the tumor necrosis factor alpha-induced and SCF-mediated degradation of IkappaB by forming catalytically inactive complexes lacking ROC1. In contrast, the C terminus of CUL1 alone interacts with ROC1 through a region containing the cullin consensus domain, to form a complex fully active in supporting ubiquitin polymerization. These results suggest the mode of action of SCF-ROC1, where CUL1 serves as a dual-function molecule that recruits an F-box protein for substrate targeting through Skp1 at its N terminus, while the C terminus of CUL1 binds ROC1 to assemble a core ubiquitin ligase.

Anaphase-Promoting Complex-Cyclosome↗
PubMed2000

Variability in CD34+ cell counts in umbilical cord blood: implications for cord blood transplants.

OBJECTIVE: To determine if total nucleated cell counts alone are sufficient for predicting the efficacy of cord blood units for transplant from neonatal umbilical cord blood samples. METHODS: Umbilical cord blood samples were collected from 200 mothers at delivery and the cord blood units processed. The total nucleated cells and CD34+ cells were enumerated and compared for each sample. RESULTS: Despite an apparent linear correlation between total nucleated cell counts and CD34+ cell counts, each group of total nucleated cell counts demonstrated a high degree of variation in CD34+ cell counts and could be as low as 0.1% of total nucleated cell counts. CONCLUSIONS: Large variations in CD34+ cell counts per total nucleated cell count are present for cord blood units from neonatal umbilical cord samples. Hence a CD34+ cell count for each cord blood unit would improve selection of samples for transplant.

Antigens, CD34↗
PubMed2000

Pseudomembranous tracheobronchitis caused by Aspergillus in a patient after peripheral blood stem cell transplantation.

INTRODUCTION: We report a case of pseudomembranous tracheobronchitis caused by Aspergillus fumigatus 2 years after matched unrelated stem cell transplant. CLINICAL PICTURE: The patient presented with dyspnoea and obstructive airway disease coinciding with the onset of chronic graft-versus-host disease (GVHD). Following treatment with higher immunosuppressive therapy for presumptive diagnosis of bronchiolitis obliterans, he subsequently developed recurrent spontaneous pneumomediastinum and progressive respiratory failure. TREATMENT AND OUTCOME: Tracheobronchial biopsy and culture of bronchoalveolar lavage (BAL) fluid revealed Aspergillus tracheobronchitis. Despite mechanical ventilation and antifungal therapy, he succumbed to progressive respiratory failure. CONCLUSIONS AND CLINICAL IMPLICATIONS: Aspergillus tracheobronchitis should be suspected in heavily immunosuppressed stem cell transplant recipients presenting with recurrent pneumomediastinum and progressive respiratory failure.

Adult↗
PubMed2000

[Clinical study on treatment of 40 cases of malignant brain tumor by elemene emulsion injection].

OBJECTIVE: To investigate the effect of elemene emulsion injection (EEI) in treating malignant brain tumor. METHODS: By conducting a retrospective study of 40 patients with brain tumor, 29 of malignant glioma and 11 metastatic tumor, who were treated with EEI from January 1994 to May 1998. EEI 0.4-1.2 g/d was given to each patient by intravenous dripping or/and intravenous infusion by pumps, and directly injected into carotid artery or infused through a carotid artery catheter with pumps. The total dosage of 6-12 g was given in 2-6 therapeutic courses with an interval of 1-1.5 months between courses. The effectiveness of treatment was accessed according to the changes of tumor size, Karnofsky Performance Status (KPS) and survival time of patients. The control group consisted of 29 cases of malignant brain tumor (22 of primary and 7 of metastatic) was treated with chemotherapy 2-3 therapeutic courses with an interval of 1-1.5 months between them. RESULTS: (1) In the EEI treated group the mean tumor size was changed from 6.70 cm3 (before treatment) to 2.67 cm3 (after treatment), t = 3.02, P < 0.01, it was reduced by 61%; (2) In the EEI treated group 4 cases was CR, 26 PR, the total effective rate being 75.0% (95% credibility interval +/- 13.4%), while in the control group, 2 of CR, 10 PR, and the total effective rate 41.4% (95% credibility interval +/- 17.9%), the difference between the two groups was significant, chi 2 = 3.867, P < 0.05; (3) KPS decreased in the EEI group from 94.7 scores (before treatment) to 88.2 scores (after treatment), the decrement was 6.5 scores (t = 3.5313, P < 0.01); (4) The survival time in the EEI treated group was 25.4 months, and that in the control group was 17.4 months (t = 3.74, P < 0.01). CONCLUSION: Elemene has significant effect on treatment of malignant brain tumor. It could prolong the high quality survival time of patients and is worthy of further investigation.

Adult↗
PubMed1999

Identification of amino acid residues critical for aggregation of human CC chemokines macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES. Characterization of active disaggregated chemokine variants.

Human CC chemokines macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES (regulated on activation normal T cell expressed) self-associate to form high-molecular mass aggregates. To explore the biological significance of chemokine aggregation, nonaggregating variants were sought. The phenotypes of 105 hMIP-1alpha variants generated by systematic mutagenesis and expression in yeast were determined. hMIP-1alpha residues Asp26 and Glu66 were critical to the self-association process. Substitution at either residue resulted in the formation of essentially homogenous tetramers at 0.5 mg/ml. Substitution of identical or analogous residues in homologous positions in both hMIP-1beta and RANTES demonstrated that they were also critical to aggregation. Our analysis suggests that a single charged residue at either position 26 or 66 is insufficient to support extensive aggregation and that two charged residues must be present. Solution of the three-dimensional NMR structure of hMIP-1alpha has enabled comparison of these residues in hMIP-1beta and RANTES. Aggregated and disaggregated forms of hMIP-1alpha, hMIP-1beta, and RANTES generally have equivalent G-protein-coupled receptor-mediated biological potencies. We have therefore generated novel reagents to evaluate the role of hMIP-1alpha, hMIP-1beta, and RANTES aggregation in vitro and in vivo. The disaggregated chemokines retained their human immunodeficiency virus (HIV) inhibitory activities. Surprisingly, high concentrations of RANTES, but not disaggregated RANTES variants, enhanced infection of cells by both M- and T-tropic HIV isolates/strains. This observation has important implications for potential therapeutic uses of chemokines implying that disaggregated forms may be necessary for safe clinical investigation.

Amino Acid Sequence↗
PubMed1999

Finding bile duct injuries using record linkage: a validated study of complications following cholecystectomy.

Laparoscopic cholecystectomy was introduced to Western Australia in 1991 and has become the method of choice for this procedure, although there are concerns about complications, particularly bile duct injuries. Previous studies have investigated this problem but have not confirmed the accuracy of coded information. We used Record Linkage to link operative admissions to subsequent admissions for all people who underwent cholecystectomy between 1988 and 1994. Using ICD9-CM discharge codes, we identified patients with an associated complication. We validated these patients' medical notes to obtain the proportion of complications in the period encompassing the introduction of laparoscopic cholecystectomy. We found 48 bile duct injuries in 413 patients. Of these 43% were found using complication codes on the operative admission, 79% using linked records of subsequent admissions, and 90% by adding lists of complicated cases from the three teaching hospitals. Any epidemiological research that uses surgical complication codes from operative admissions, particularly in the absence of a specific ICD9-CM code, will lead to significantly underestimating the prevalence of complications. By using record linkage, and validating medical records, we captured a significant proportion of complications.

Bile Ducts↗
PubMed1999

Homozygosity for a nonsense mutation in the alpha-tropomyosin slow gene TPM3 in a patient with severe infantile nemaline myopathy.

The nemaline myopathies are muscle disorders of variable severity and age of onset, with characteristic nemaline bodies in the sarcoplasm. Genes for dominant (NEM1) and recessive (NEM2A) nemaline myopathy have been localised to chromosomes one and two, respectively. A missense mutation in the alpha-tropomyosin gene (TPM3) has been associated with NEM1 in one family. Probands from 76 other nemaline myopathy families have now been screened for TPM3 mutations. One proband, who was not noted to have any weakness neonatally, but who died at 21 months of age, was shown to be homozygous for a single strand conformation polymorphism (SSCP) in skeletal-muscle-specific exon 1 of TPM3. Sequencing revealed homozygosity for a nonsense mutation at codon 31 (CAG to TAG). The patient should have no functioning alpha-tropomyosin slow protein. The nemaline bodies in this patient were exclusively in type one fibres, consistent with the expression of TPM3 only in type one fibres.

Fatal Outcome↗
PubMed1999

Recruitment of a ROC1-CUL1 ubiquitin ligase by Skp1 and HOS to catalyze the ubiquitination of I kappa B alpha.

Activation of the transcription factor NF-kappa B in response to proinflammatory stimuli requires the phosphorylation-triggered and ubiquitin-dependent degradation of the NF-kappa B inhibitor, I kappa B alpha. Here, we show the in vitro reconstitution of the phosphorylation-dependent ubiquitination of I kappa B alpha with purified components. ROC1, a novel SCF-associated protein, is recruited by cullin 1 to form a quatemary SCFHOS-ROC1 holenzyme (with Skp1 and the beta-TRCP homolog HOS). SCFHOS-ROC1 binds IKK beta-phosphorylated I kappa B alpha and catalyzes its ubiquitination in the presence of ubiquitin, E1, and Cdc34. ROC1 plays a unique role in the ubiquitination reaction by heterodimerizing with cullin 1 to catalyze ubiquitin polymerization.

Amino Acid Sequence↗
PubMed1999

Cellular and molecular mechanisms of inflammation and thrombosis.

In the last 20 years, the cellular and molecular mechanisms of inflammation and thrombosis have been characterised. These are essentially cell adhesion processes which are regulated by vascular endothelium. Many of the cell adhesion molecules and leucocyte chemoattractants expressed and generated at sites of inflammation have been sequenced and cloned. These inflammatory molecules work together in concert to mediate the adhesion between leucocytes, platelets and vascular endothelium which occurs during the occlusive, thromboembolic, reperfusion and septic complications of atherosclerotic and diabetic vascular diseases. This review aims to summarise our current understanding of the molecular basis of these disorders and the therapeutic implications.

Animals↗
PubMed1999

Complications of cholecystectomy: risks of the laparoscopic approach and protective effects of operative cholangiography: a population-based study.

BACKGROUND: Previous studies suggest that laparoscopic cholecystectomy (LC) is associated with an increased risk of intraoperative injury involving the bile ducts, bowel, and vascular structures compared with open cholecystectomy (OC). Population-based studies are required to estimate the magnitude of the increased risk, to determine whether this is changing over time, and to identify ways by which this might be reduced. METHODS: Suspected cases of intraoperative injury associated with cholecystectomy in Western Australia in the period 1988 to 1994 were identified from routinely collected hospital statistical records and lists of persons undergoing postoperative endoscopic retrograde cholangiopancreatography. The case records of suspect cases were reviewed to confirm the nature and site of injury. Ordinal logistic regression was used to estimate the risk of injury associated with LC compared with OC after adjusting for confounding factors. RESULTS: After the introduction of LC in 1991, the proportion of all cholecystectomy cases with intraoperative injury increased from 0.67% in 1988-90 to 1.33% in 1993-94. Similar relative increases were observed in bile duct injuries, major bile leaks, and other injuries to bowel or vascular structures. Increases in intraoperative injury were observed in both LC and OC. After adjustment for age, gender, hospital type, severity of disease, intraoperative cholangiography, and calendar period, the odds ratio for intraoperative injury in LC compared with OC was 1.79. Operative cholangiography significantly reduced the risk of injury. CONCLUSION: Operative cholangiography has a protective effect for complications of cholecystectomy. Compared with OC, LC carries a nearly twofold higher risk of major bile, vascular, and bowel complications. Further study is required to determine the extent to which potentially preventable factors contribute to this risk.

Aged↗