Biomedical subjects

G Lee

Publications and source records attributed to G Lee.

At least 37 records · Page 2Linked to original sources

PubMed2003

Cell cycle aberrations by alpha-synuclein over-expression and cyclin B immunoreactivity in Lewy bodies.

alpha-Synuclein is a presynaptic protein that accumulates abnormally in Lewy bodies of Parkinson's disease (PD) and dementia with Lewy bodies (DLB). Its physiological function and role in neuronal death remain poorly understood. Recent immunohistochemical studies suggest that cell cycle-related phenomena may play a role in the pathogenesis of Alzheimer's disease and perhaps other neurodegenerative disorders. In this investigation, we examined the effects of alpha-synuclein expression levels on cell cycle indices in PC12 cells engineered to conditionally induce alpha-synuclein expression upon withdrawal of doxycycline. Over-expression of alpha-synuclein resulted in enhanced proliferation rate and enrichment of cells in the S phase of the cell cycle. This was associated with increased accumulation of the mitotic factor cyclin B and down-regulation of the tumor suppressor retinoblastoma 2. Additionally, ERK1/2, key molecules in proliferation signaling, were highly phosphorylated. Immunohistochemical studies on postmortem brains revealed intense cyclin B immunoreactivity in Lewy bodies in cases with DLB and to a lesser extent in PD. We propose that elevated expression of alpha-synuclein causes changes in cell cycle regulators through ERK activation leading to apoptosis of postmitotic neurons. These changes in cell cycle proteins are also associated with ectopic expression of cyclin B in Lewy bodies.

Animals↗

PubMed2003

Recurrent haemorrhage from a malignant brain stem glioma: a review of the mechanisms of tumoural haemorrhage.

The authors report a 23-year-old man who was diagnosed with a brain stem malignant glioma following his first episode of generalised seizure. This was subsequently complicated by three separate documented episodes of intracranial haemorrhage. The literature is reviewed and the pathophysiological mechanisms of tumoural haemorrhage are discussed. It is speculated that significant intratumoural arteriovenous shunting may lead to recurrent haemorrhages.

Brain Stem Neoplasms↗

PubMed2002

Goosebumps.

Explore the source record for details and available documents.

Astrocytoma↗

PubMed2002

The impact of ancillary HIV services on engagement in medical care in New York City.

The advent of antiretroviral therapies in 1996 prompted an interest in the role played by ancillary services in improving access to and retention in medical care, particularly since the success of the new therapies is often contingent upon ongoing and appropriate primary medical care. Using self-reported survey data from a longitudinal representative sample of 577 HIV-positive adults in New York City, this paper explores the impact of such supportive services as drug treatment, case management, housing assistance, mental health treatment and transportation on engagement with medical care. The study's principal finding was that specific ancillary services were significantly associated with an increase in an individual's likelihood of entering medical care and maintaining appropriate medical care services for HIV, particularly when the services addressed a corresponding need.

Adult↗

PubMed2002

The effect on IVF outcome of small intramural fibroids not compressing the uterine cavity as determined by a prospective matched control study.

BACKGROUND: Several studies have reported that the presence of intramural fibroids affects conception following IVF. We attempted to corroborate or refute the conclusions relating to IVF and leiomyomas of the aforementioned studies. METHODS: Women with small intramural leiomyomata (< or = 5 cm) discovered on initial pelvic sonographic studies performed in preparation for IVF were prospectively matched by age, with the next patient of the same age undergoing IVF who did not demonstrate fibroids (controls). RESULTS: Though no significant differences were found in outcome when comparing these two groups, there was a distinct trend for lower live delivery rates and higher miscarriage rates. CONCLUSIONS: These data support the conclusions of the only other prospective matched control study evaluating similar factors, i.e. that small intramural fibroids can negatively affect IVF outcome. Nevertheless, we think that a multicentre study should be conducted first before evaluating whether myomectomy improves outcome or not.

Abortion, Spontaneous↗

PubMed2002

PCR detection of Mycobacterium paratuberculosis in Crohn's disease granulomas isolated by laser capture microdissection.

BACKGROUND AND AIMS: The uncertainty surrounding the role of Mycobacterium avium subsp paratuberculosis (Map) in Crohn's disease has been compounded by possible contamination from Map present in the lumen microflora. This study used laser capture microdissection (LCM) and polymerase chain reaction (PCR) to detect Map DNA in subepithelial granulomas, isolated from 15 surgically resected, formalin fixed specimens of granulomatous Crohn's disease and from 12 granulomatous disease controls (10 bowel, 2 non-bowel). METHODS: The effect of amplicon size on reliability of PCR from formalin fixed samples was examined by amplifying 435 bp and 133 bp sequences of the human APC gene. After this, nested primers were designed to detect a small fragment (155 bp) of the Map specific IS900 gene in Crohn's granulomas. LCM isolated granulomas from Map culture positive bovine intestine was used as positive control. PCR product specificity was confirmed by direct DNA sequencing. RESULTS: The smaller, but not the larger, fragment of the APC gene amplified reliably in all samples. Amplification of the 155 bp fragment of the IS900 gene detected Map DNA in microdissected Crohn's granulomas in 6 of 15 cases, and in 0 of 12 disease control granulomas. CONCLUSIONS: LCM can be used to detect Map DNA in granulomas in a proportion of patients with Crohn's disease. However, formalin fixation requires that comparatively short DNA fragments of the Map specific IS900 gene be targeted, to permit consistent detection. Detection of Map DNA within granulomas might suggest an infectious aetiology in a subset of patients; alternatively, a transmissible agent may not be involved but mycobacterial DNA may influence pathogenesis by modifying the local cytokine responses.

Case-Control Studies↗

PubMed2002

Human herpesvirus 8 (HHV-8) detected in two patients with Kaposi's sarcoma-like pyogenic granuloma.

AIMS: To report the finding of human herpesvirus 8 (HHV-8) in two patients with Kaposi's sarcoma (KS)-like pyogenic granuloma. This form of pyogenic granuloma closely resembles KS histologically and it has been reported that immunohistochemistry in such lesions may be positive for smooth muscle actin and factor VIII related antigen, which are typically negative in KS. In both patients the lesions were positive for CD31, CD34, smooth muscle actin, and factor VIII related antigen, a profile typical of KS-like pyogenic granuloma. The lesions were tested for the presence of HHV-8 DNA, which to date has been consistently found in all types of KS. METHODS: The lesions were tested for the presence of HHV-8 DNA using the polymerase chain reaction (PCR). A known HHV-8 positive KS specimen was used as the positive control. Six samples of non-KS vascular skin lesions were used as negative controls for the PCR reaction. RESULTS: Both lesions were positive on PCR for HHV-8 and the specificity of product was confirmed by direct sequencing. None of the six control vascular skin lesions was positive for HHV-8. These results strongly indicate KS as the true diagnosis and are supported by the reported clinical course in both cases. CONCLUSIONS: Techniques targeting HHV-8 DNA for detection to confirm a diagnosis of KS are both sensitive and specific. In cases where the differential diagnosis includes KS-like pyogenic granuloma, caution should be taken not to diagnose solely on the basis of immunohistochemistry phenotype. In such cases, PCR targeting HHV-8 DNA sequences is a better diagnostic tool.

Aged↗

PubMed2001

Alternative transcriptional initiation and splicing of mouse Lamc2 message.

To investigate the mechanism of two forms of messages (5.1 and 2.4 kb) in mouse Lamc2, a gene encoding for the gamma2 chain of epithelial cell-specific laminin 5, we analyzed approximately 40 kb of genomic DNA containing the sequences from the first intron to the 23rd exon to the 3' untranscribed region. With the 5' rapid amplification of cDNA end (RACE) and primer extension technique using RNA from mouse kidney and thymic epithelial cells, we found that the smaller message starts at the 2895th base of the cDNA within the 19th exon. Upstream sequences of this alternative start site showed the promoter activity in the reporter assay with the secreted form of alkaline phosphatase (SEAP). We also identified a novel alternatively spliced exon (exon 19B) that includes two stop codons. These results revealed two possible additional open reading frame that are different from the previously described alternative human gamma2 peptide. Therefore, the mechanism for generating smaller message and the event of alternative splicing are quite distinct between mouse and human Lamc2, although genomic organization is highly conserved. The significance of the presence of alternative message is discussed.

Alternative Splicing↗

PubMed2001

Analysis of integral membrane protein contributions to the deformability and stability of the human erythrocyte membrane.

Three major hypotheses have been proposed to explain the role of membrane-spanning proteins in establishing/maintaining membrane stability. These hypotheses ascribe the essential contribution of integral membrane proteins to (i) their ability to anchor the membrane skeleton to the lipid bilayer, (ii) their capacity to bind and stabilize membrane lipids, and (iii) their ability to influence and regulate local membrane curvature. In an effort to test these hypotheses in greater detail, we have modified both the membrane skeletal and lipid binding interactions of band 3 (the major membrane-spanning and skeletal binding protein of the human erythrocyte membrane) and have examined the impact of these modifications on erythrocyte membrane morphology, deformability, and stability. The desired changes in membrane skeletal and protein-lipid interactions were induced by 1) reaction of the cells with 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS), an inhibitor of band 3-mediated anion transport that dissociates band 3 into dimers (increasing its surface area in contact with lipid) and severs band 3 linkages to the membrane skeleton; 2) a fragment of ankyrin that ruptures the same ankyrin-band 3 bridge to the membrane skeleton, but drives the band 3 subunit equilibrium toward the tetramer (i.e. decreasing the band 3 surface area in contact with lipid); and 3) an antibody to the ankyrin-binding site on band 3 that promotes the same changes in band 3 skeletal and lipid interactions as the ankyrin fragment. We observed that although DIDS induced echinocytic morphological changes in the treated erythrocytes, it had little impact on either membrane deformability or stability. In contrast, resealing of either the ankyrin fragment or anti-band 3 IgG into erythrocytes caused spontaneous membrane fragmentation and loss of deformability/stability. Because these and other new observations cannot all be reconciled with any single hypothesis on membrane stability, we suggest that more than one hypothesis may be operative and provide an explanation of how each might individually contribute to net membrane stability.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed2001

An unusually low pK(a) for Cys282 in the active site of human muscle creatine kinase.

All phosphagen kinases contain a conserved cysteine residue which has been shown by crystallographic studies, on both creatine kinase and arginine kinase, to be located in the active site. There are conflicting reports as to whether this cysteine is essential for catalysis. In this study we have used site-directed mutagenesis to replace Cys282 of human muscle creatine kinase with serine and methionine. In addition, we have replaced Cys282, conserved across all creatine kinases, with alanine. No activity was found with the C282M mutant. The C282S mutant showed significant, albeit greatly reduced, activity in both the forward (creatine phosphorylation) and reverse (MgADP phosphorylation) reactions. The K(m) for creatine was increased approximately 10-fold, but the K(m) for phosphocreatine was relatively unaffected. The V and V/K pH-profiles for the wild-type enzyme were similar to those reported for rabbit muscle creatine kinase, the most widely studied creatine kinase isozyme. However, the V/K(creatine) profile for the C282S mutant was missing a pK of 5.4. This suggests that Cys282 exists as the thiolate anion, and is necessary for the optimal binding of creatine. The low pK of Cys282 was also determined spectrophotometrically and found to be 5.6 +/- 0.1. The S284A mutant was found to have reduced catalytic activity, as well as a 15-fold increase in K(m) for creatine. The pK(a) of Cys282 in this mutant was found to be 6.7 +/- 0.1, indicating that H-bonding to Ser284 is an important, but not the sole, factor contributing to the unusually low pK(a) of Cys282.

Amino Acid Sequence↗

PubMed2001

Use of vascularized fibular grafts as a salvage procedure for previously failed spinal arthrodesis.

STUDY DESIGN: A case series of spinal arthrodesis performed with vascularized fibular grafts is presented. OBJECTIVES: The use of vascularized fibular grafts to obtain anterior spinal fusion in patients with complex spinal disorders and poor fusion environments is described. SUMMARY OF BACKGROUND DATA: The fusion success of spinal arthrodesis is dependent on numerous factors, such as the substrate used for arthrodesis (i.e., graft), the biology of the fusion bed, and local host factors. Vascularized grafts have higher success rates for union and can better overcome a poor fusion bed than nonvascular grafts. However, they are associated with higher donor site morbidity and greater technical difficulty. METHODS: Three patients with complex medical histories portending a difficult spinal fusion were treated with anterior arthrodesis using vascularized fibular autografts. Vascular patency was confirmed by bone scintigraphy, and osseous union by radiography and computed tomography. RESULTS: All patients had successful osseous fusion in 3 to 6 months. Deformity was improved. Patients reported decreased pain and resumption to previous activities at 24 months follow-up. There were no complications. CONCLUSIONS: A vascularized fibular graft is a useful alternative to standard grafts for spinal arthrodesis. Vascularized fibular grafts provide high fusion rates, rapid incorporation, and increased mechanical strength, and thus heal better in a suboptimal graft bed. The procedure is technically demanding, often requiring the expertise of a microvascular surgeon for obtaining the graft and achieving anastomosis. It is best suited in cases where significant difficulty in obtaining a spinal fusion is anticipated.

Adult↗

PubMed2001

Binding of Fyn to MAP-2c through an SH3 binding domain. Regulation of the interaction by ERK2.

Microtubule-associated protein 2 (MAP-2) isoforms are developmentally expressed in the nervous system and contain a number of functional domains. Adjacent to the first repeat of the microtubule-binding domain is an RTPPKSP motif for binding SH3 domains. To identify SH3-containing proteins that interact with MAP-2, transfections, filter overlay assays, glutathione S-transferase (GST)-mediated binding assays, co-immunoprecipitations and enzyme-linked immunosorbent assays were performed. Transfections of MAP-2a, MAP-2b, and MAP-2c constructs into COS7 cells, followed by incubation of the cell lysates with SH3-GST fusion proteins, determined that the strongest interaction was between MAP-2c and the non-receptor tyrosine kinase Fyn; however, MAP-2b and MAP-2c also bound to Grb2. Co-immunoprecipitation of Fyn and MAP-2c from human fetal homogenates confirmed the interaction in vivo. MAP-2 synthetic peptides spanning the RTPPKSP motif bound to Fyn, and the interaction was regulated by phosphorylation. Co-transfections with MAP-2c and the extracellular signal-regulated kinase 2 (ERK2) demonstrated that MAP-2c is threonine/serine-phosphorylated on its RTPPKSP motif and that threonine phosphorylation abolished the MAP-2c/Fyn binding. Kinase assays and co-transfection of MAP-2c and Fyn confirmed that Fyn tyrosine kinase phosphorylates MAP-2c. Thus, the activation of signaling pathways may regulate cytoskeletal dynamics by altering the state of phosphorylation of MAP-2 by both ERK2 and Fyn kinase.

Adaptor Proteins, Signal Transducing↗

PubMed2001

Structure of the Ba-induced Si(111)- (3 x 2) reconstruction.

The Ba/Si(111) surface, previously known as a 3 x 1 phase, is found to have a 3 x 2 periodicity and a semiconducting band gap. The substrate reconstructs into the honeycomb chain-channel (HCC) structure with Ba atoms in the channel, as in the alkali-metal-induced Si(111)-(3 x 1). However, the metal coverage is determined to be 1/6 monolayers, half the alkali-metal coverage. We propose that the structure and the metal coverage determined for the Ba adsorbate is universal for other alkaline-earth-metal adsorbates. With the alkali-metal-induced 3 x 1 case, our results lead to a rule that one donated electron per 3 x 1 surface unit is necessary to stabilize the HCC reconstruction of Si.

Journal Article↗

PubMed2001

Glycophorin A dimerization and band 3 interaction during erythroid membrane biogenesis: in vivo studies in human glycophorin A transgenic mice.

Band 3 and glycophorin A (GPA) are the 2 most abundant integral proteins in the human erythrocyte membrane. Earlier studies suggested that the 2 proteins may associate not only in the mature erythrocyte membrane, but also during their posttranslational processing and intracellular trafficking. The purpose of this study was to directly examine the GPA-band 3 interaction in vivo and determine the nature of this association during erythroid membrane biogenesis. Transgenic mice were generated expressing the human glycophorin A gene and were used to examine how the induction of human GPA expression affected the levels of murine GPA and band 3 expression in the red cell membrane. Murine GPA expression was reduced in erythrocytes expressing human GPA, whereas the level of band 3 expression remained constant, implying a tight coupling of band 3 and GPA expression in the membrane of mature red cells. In vivo GPA dimerization was not modulated solely by the GPA transmembrane motif, but the distance between this motif and the basic residues on the cytoplasmic side of the transmembrane domain may also be important. In addition, GPA monomers with varying degrees of glycosylation dimerized, providing clear evidence that carbohydrate structures on the extracellular domain do not affect dimerization. The association between the multiple transmembrane-spanning protein, band 3, and the single transmembrane-spanning sialoglycoprotein, GPA, may serve as a model for interactions of other multi-pass and single-pass polypeptides during membrane biogenesis.

Animals↗

PubMed2001

Evaluation of a rapid qualitative prostate specific antigen assay, the One Step PSA(TM) test.

Recently, highly sensitive prostate specific antigen (PSA) kits have been developed and reported to be useful for the early identification of a chemical relapse. However, if the measurement time was short and the cost low, such an assay kit should be sufficient for cancer screening when dealing with a large number of samples. The One Step PSA test uses an immunochromatographic method to qualitatively, not quantitatively, judge a positive or negative result. We confirmed the sensitivity of the kit using purified PSA. Serum specimens from 147 men with or without prostate diseases were tested using the kit. PSA concentration of each serum specimen was independently measured by a quantitative ACS-PSA2 EIA kit (Chiron, cut-off: 2.1 ng/ml). The sensitivity of this kit was determined to be 4 ng/ml. All 33 samples with a value of greater than 4 ng/ml were clearly positive. Of the 94 samples with values less than 4 ng/ml, nine were judged as positive. The remaining 85 cases were judged as completely negative. These results indicate that the sensitivity of the One Step PSA test is 100% and the specificity is 90.4%. Tests using this kit can be easily performed at outpatient clinics or elsewhere. This kit is useful for initial cancer screening, because results can be obtained within 15 min and at a cost lower than that of ordinary PSA kits.

Humans↗

PubMed2001

Abnormalities of male-specific FRU protein and serotonin expression in the CNS of fruitless mutants in Drosophila.

The fruitless gene in Drosophila produces male-specific protein (FRU(M)) involved in the control of courtship. FRU(M) spatial and temporal patterns were examined in fru mutants that exhibit aberrant male courtship. Chromosome breakpoints at the locus eliminated FRU(M). Homozygous viable mutants exhibited an intriguing array of defects. In fru(1) males, there were absences of FRU(M)-expressing neuronal clusters or stained cells within certain clusters, reductions of signal intensities in others, and ectopic FRU(M) expression in novel cells. fru(2) males exhibited an overall decrement of FRU(M) expression in all neurons normally expressing the gene. fru(4) and fru(sat) mutants only produced FRU(M) in small numbers of neurons at extremely low levels, and no FRU(M) signals were detected in fru(3) males. This array of abnormalities was inferred to correlate with the varying behavioral defects exhibited by these mutants. Such abnormalities include courtship among males, which has been hypothesized to involve anomalies of serotonin (5-HT) function in the brain. However, double-labeling uncovered no coexpression of FRU(M) and 5-HT in brain neurons. Yet, a newly identified set of sexually dimorphic FRU(M)/5-HT-positive neurons was identified in the abdominal ganglion of adult males. These sexually dimorphic neurons (s-Abg) project toward regions of the abdomen involved in male reproduction. The s-Abg neurons and the proximal extents of their axons were unstained or absent in wild-type females and exhibited subnormal or no 5-HT immunoreactivity in certain fru-mutant males, indicating that fruitless controls the formation of these cells or 5-HT production in them.

Animals↗

PubMed2001

Lutheran blood group glycoprotein and its newly characterized mouse homologue specifically bind alpha5 chain-containing human laminin with high affinity.

Lutheran blood group glycoproteins (Lu gps) are receptors for the extracellular matrix protein, laminin. Studies suggest that Lu gps may contribute to vaso-occlusion in sickle cell disease and it has recently been shown that sickle cells adhere to laminin isoforms containing the alpha5 chain (laminin 10/11). Laminin alpha5 is present in the subendothelium and is also a constituent of bone marrow sinusoids, suggesting a role for the Lu/laminin interaction in erythropoiesis. The objectives of the current study were to define more precisely the molecular interactions of the extracellular and intracellular regions of human Lu and to clone and characterize a mouse homologue. To this end, complementary DNA and genomic clones for the mouse homologue were sequenced and the mouse Lu gene mapped to a region on chromosome 7 with conserved synteny with human 19q13.2. Mouse and human Lu gps are highly conserved (72% identity) at the amino acid sequence level and both mouse and human Lu gps specifically bind laminin 10/11 with high affinity. Furthermore, the first 3, N-terminal, immunoglobulin superfamily domains of human Lu are critical for this interaction. The results indicated that the cytoplasmic domain of BRIC 221-labeled human Lu gp is linked with the spectrin-based skeleton, affording the speculation that this interaction may be critical for signal transduction. These results further support a role for Lu gps in sickle cell disease and indicate the utility of mouse models to explore the function of Lu gp-laminin 10/11 interaction in normal erythropoiesis and in sickle cell disease.

Amino Acid Sequence↗

PubMed2001

Continuous measurement of drying rate of crystalline and amorphous systems during freeze-drying using an in situ microbalance technique.

The use of a novel microbalance (Christ) technique to monitor continuously the weight loss of a vial standing on a shelf of a freeze-dryer has been investigated. The drying rates of the following aqueous solutions were measured during the primary drying phase of a complete freeze-drying cycle: sucrose (75 mg/mL, 2.5-mL fill volume), sucrose and phenylalanine (1:0.2 by weight, 75 mg/mL, 2.5-mL fill volume), and mannitol (75mg/mL, 2.5-mL fill volume). The microbalance yields the cumulative water loss, m(cu) in grams, and the momentary drying rate, Deltam(cu)/Deltat in mg/10 min, of the frozen cake. The momentary drying rate curves were especially useful for examining how Deltam(cu)/Deltat changes with time during primary drying. Initially, Deltam(cu)/Deltat rises to a sharp maximum and then decreases in a fashion depending on shelf temperature, chamber pressure, and the nature of the substance being dried. Different drying behavior was observed for the sucrose and sucrose/phenylalanine systems, which was attributed to the presence of crystalline phenylalanine in the amorphous sucrose. At low shelf-temperature (-24 degrees C) the crystalline mannitol showed lower Deltam(cu)/Deltat than with either sucrose or sucrose/phenylalanine. The balance could also detect differences in Deltam(cu)/Deltat when using different freezing protocols. "Slow" and "moderate" freezing protocols gave similar drying behavior, but "rapid" freezing in liquid nitrogen produced greatly altered drying rate and internal cake morphology. The balance also could be used to detect the endpoint of primary drying. Different endpoint criteria and their influence on final dried cake properties were examined.

Crystallization↗