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J Lim

Publications and source records attributed to J Lim.

At least 37 records · Page 2Linked to original sources
PubMed1999

Human basophils express CD22 without expression of CD19.

BACKGROUND: Even modern automatic cell counters cannot count basophils precisely. Therefore, we need a rapid, accurate, precise, and easy method for counting basophils. METHODS: Using flow cytometry, basophils (CD22+/CD19-) and B cells (CD22+/CD19+) were counted. Within a large lymphocyte light scatter gate, % basophils (G%baso) and % B cells (G%B) were determined from the total count. Another method of analysis was to make two regions (R1 for basophils and R2 for B cells) and to determine in those the % basophils (R1%baso) and % B cells (R2%B) without gating. The flow cytometric basophil counts of the blood of 21 normal controls and 43 chronic myelogenous leukemia (CML) patients were compared with manual basophil count (Ma%baso) and basophil count by Coulter electronic cell counter (Hialeah, FL) (Auto%baso). CD22+/CD19- cells were sorted by a FACSCalibur (Becton Dickinson, San Jose, CA). RESULTS: The G%baso of all samples was 4.66 +/- 5.35%, and R1%baso was 4.23 +/- 4.88%, and they were well-correlated (r = 0.996, P < 0.001). The G%B of all samples was 1.55 +/- 1.68%, and R2%B was 1.59 +/- 1.67%, and they were also well-correlated (r = 0.993, P < 0.001). Their correlation was better in normal controls than in CML. G%baso was well-correlated to Ma%baso (r = 0.827) and Auto%baso (r = 0.806), and R1%baso was well-correlated to Ma%baso (r = 0.831) but showed poor correlation to Auto%baso (r = 0.734). Auto%baso revealed the poorest correlation to Ma%baso (r = 0.692). The sorted CD22+/CD19- cells were all basophils (99.48 +/- 0.30%), and they revealed CD13, CD33, and dim CD45 expression, whereas CD3, CD14, CD16, and HLA-DR were not detected on them. CONCLUSIONS: We discovered a specific marker combination to identify basophils (CD22+/CD19-), and we suggest that flow cytometric analysis using these markers is an easy, reliable, and accurate method of basophil counting.

Antigens, CD↗
PubMed1999

Activation of the cAMP-specific phosphodiesterase PDE4D3 by phosphorylation. Identification and function of an inhibitory domain.

Splicing variants of type 4 phosphodiesterases (PDE4) are regulated by phosphorylation. In these proteins, a conserved region is located between the amino-terminal domain, which is the target for phosphorylation, and the catalytic domain. Previous studies have indicated that nested deletions encompassing this region cause an increase in catalytic activity, suggesting this domain exerts an inhibitory constraint on catalysis. Here, we have further investigated the presence and function of this domain. A time-dependent increase in hydrolytic activity was observed when PDE4D3 from FRTL-5 cells was incubated with the endoproteinase Lys-C. The activation was abolished by protease inhibitors and was absent when a phosphorylated enzyme was used. Western blot analysis with PDE4D-specific antibodies indicated the Lys-C treatment separates the catalytic domain of PDE4D3 from the inhibitory domain. Incubation with antibodies recognizing an epitope within this domain caused a 3- to 4-fold increase in activity of native or recombinant PDE4D3. Again, PDE activation by these antibodies had properties similar to, and not additive with, the activation by protein kinase A phosphorylation. An interaction between the inhibitory domain and both regulatory and catalytic domains of PDE4D3 was detected by the yeast two-hybrid system. Mutations of Ser54 to Ala in the regulatory domain decreased or abolished this interaction, whereas mutations of Ser54 to the negatively charged Asp strengthened it. These data strongly support the hypothesis that an inhibitory domain is present in PDE4D and that phosphorylation of the regulatory domain causes activation of the enzyme by modulating the interaction between inhibitory and catalytic domains.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1999

Association of atypical protein kinase C isotypes with the docker protein FRS2 in fibroblast growth factor signaling.

FRS2 is a docker protein that recruits signaling proteins to the plasma membrane in fibroblast growth factor signal transduction. We report here that FRS2 was associated with PKC lambda when Swiss 3T3 cells were stimulated with basic fibroblast growth factor. PKC zeta, the other member of the atypical PKC subfamily, could also bind FRS2. The association between FRS2 and PKC lambda is likely to be direct as shown by yeast two-hybrid analysis. The C-terminal fragments of FRS2 (amino acid residues 300-508) and SNT2 (amino acids 281-492), an isoform bearing 50% identity to FRS2, interacted with PKC lambda at a region (amino acids 240-562) that encompasses the catalytic domain. In vitro kinase assays revealed neither FRS2 nor SNT2 was a substrate of PKC lambda or zeta. Mutation of the alanine residue (Ala-120) to glutamate in the pseudo-substrate region of PKC lambda results in a constitutively active kinase that exhibited more than 2-fold greater binding to FRS2 in vitro than its "closed" wild-type counterpart. Tyrosine phosphorylation of FRS2 did not affect its binding to the constitutively active PKC lambda mutant, suggesting that the activation of PKC lambda is necessary and sufficient for its association with FRS2. It is likely that FRS2 serves as an anchoring protein for targeting activated atypical PKCs to the cell plasma membrane in signaling pathways.

3T3 Cells↗
PubMed1999

The A beta peptide of Alzheimer's disease directly produces hydrogen peroxide through metal ion reduction.

Oxidative stress markers characterize the neuropathology both of Alzheimer's disease and of amyloid-bearing transgenic mice. The neurotoxicity of amyloid A beta peptides has been linked to peroxide generation in cell cultures by an unknown mechanism. We now show that human A beta directly produces hydrogen peroxide (H2O2) by a mechanism that involves the reduction of metal ions, Fe(III) or Cu(II), setting up conditions for Fenton-type chemistry. Spectrophotometric experiments establish that the A beta peptide reduces Fe(III) and Cu(II) to Fe(II) and Cu(I), respectively. Spectrochemical techniques are used to show that molecular oxygen is then trapped by A beta and reduced to H2O2 in a reaction that is driven by substoichiometric amounts of Fe(II) or Cu(I). In the presence of Cu(II) or Fe(III), A beta produces a positive thiobarbituric-reactive substance (TBARS) assay, compatible with the generation of the hydroxyl radical (OH.). The amounts of both reduced metal and TBARS reactivity are greatest when generated by A beta 1-42 >> A beta 1-40 > rat A beta 1-40, a chemical relationship that correlates with the participation of the native peptides in amyloid pathology. These findings indicate that the accumulation of A beta could be a direct source of oxidative stress in Alzheimer's disease.

Alzheimer Disease↗
PubMed1999

Accounting for differences in dieting status: steps in the refinement of a model.

OBJECTIVE: The overriding objective of this paper is to outline the steps involved in refining a structural model to explain differences in dieting status. METHODS: Cross-sectional data (representing the responses of 1,644 teenage girls) derive from the preliminary testing in a 3-year longitudinal study. A battery of measures assessed social influence, vulnerability (to conformity) disposition, protective (social coping) skills, and aspects of positive familial context as core components in a model proposed to account for the initiation of dieting. Path analyses were used to establish the predictive ability of those separate components and their interrelationships in accounting for differences in dieting status. RESULTS: Several components of the model were found to be important predictors of dieting status. The model incorporates significant direct, indirect (or mediated), and moderating relationships. Taking all variables into account, the strongest prediction of dieting status was from peer competitiveness, using a new scale developed specifically for this study. CONCLUSION: Systematic analyses are crucial for the refinement of models to be used in large-scale multivariate studies. In the short term, the model investigated in this study has been shown to be useful in accounting for cross-sectional differences in dieting status. The refined model will be most powerfully employed in large-scale time-extended studies of the initiation of dieting to lose weight.

Adaptation, Psychological↗
PubMed1999

Improved device definition in interventional magnetic resonance imaging using a rotated stripes keyhole acquisition.

Keyhole acquisition techniques have been used to reduce image acquisition times primarily in contrast agent studies and via simulation in interventional MRI procedures. More recent simulations have suggested that improved definition of an interventional device [e.g., biopsy needles, radio frequency (RF) electrodes] could be achieved by rotating the keyhole pattern in k-space so that the read out direction lies perpendicular to the device orientation in real space. This study seeks to validate the earlier predictions of improved efficacy of a rotated stripes keyhole acquisition in actual in vitro and in vivo interventional MR imaging procedures. A true-FISP sequence was modified to perform central stripes keyhole (as known as conventional keyhole) acquisitions after a full initial reference data set was acquired. The gradients of this sequence were then modified to rotate the k-space definition and the keyhole stripes by 10 degrees, 20 degrees, 30 degrees, 45 degrees, and 60 degrees from their conventional k-space orientation. Acquisitions were performed during insertion of interventional devices in phantom and in vivo RF ablation procedures, using the modified sequence selected which placed the phase encoding axis at parallel and perpendicular orientations to the devices. Resulting images were compared between the two orientations for needle width and tip accuracy. Apparent needle width was thinner and tip position more accurately determined for placement of phase encoding parallel to the needle in all cases. Rotated keyhole imaging provides the required temporal advantage of conventional keyhole imaging along with a near optimal definition of an interventional device when the phase encoding is oriented parallel to the direction of the needle motion. Magn Reson Med 42:554-560, 1999.

Animals↗
PubMed1999

Endometrial pattern on the day of oocyte retrieval is more predictive of implantation success than the pattern or thickness on the day of hCG administration.

PURPOSE: Multiple studies have confirmed a lower implantation (IR) and pregnancy rate (PR) in women who exhibit a homogeneous pattern (pattern II) of the endometrium compared to a triple-line pattern (pattern I) on the day of hCG administration. However, no data are available to evaluate if patients alter their endometrial thickness and pattern between the day of hCG administration (DhCG) and the day of oocyte retrieval (DRET) and whether these changes adversely affect endometrial receptivity. METHODS: We prospectively evaluated 86 women (mean age, 32.9 +/- 3.8 years; range, 24-40 years) undergoing 103 IVF/ET cycles. RESULTS: Pattern II was noted in 7 cycles (6.8%) on DhCG, compared to 96 cycles with pattern I (93.2%). However, 20 cycles (19.4%) had pattern II on DRET. The ongoing IR was 13.0% (3/23) in the pattern II group compared to 20.8% (76/365) in the pattern I group on DhCG (P = NS). However, a significant decrease in the ongoing IR, to 9.9% (7/71), was noted in pattern II, compared to 23.3% (71/305) in pattern I, on DRET (P = 0.019). There was no difference in age, basal FSH, peak E2, P4 on the day of hCG, number of oocytes, number of ET, or endometrial thickness between pregnant and nonpregnant patients, or between patients with pattern I and those with pattern II. A trend toward higher progesterone levels on DhCG was noted in women with pattern II (P = 0.078). CONCLUSIONS: Endometrial pattern, rather than thickness, on the day of oocyte retrieval appears to be an important prognosticator of endometrial receptivity.

Adult↗
PubMed1999

The effect of pituitary desensitization on ovarian volume measurements prior to in-vitro fertilization.

The measurement of ovarian volume has been recently shown to predict follicular response in in-vitro fertilization (IVF), specifically a lower number of retrieved oocytes with decreasing ovarian volume. This test appears to be better than basal follicle-stimulating hormone (FSH) as a prognostic measure of ovarian reserve. However, the effect of pituitary desensitization on ovarian volume has not been previously investigated. We prospectively evaluated 38 women undergoing IVF using a long luteal leuprolide acetate (LA) protocol. All women had their ovarian volume measurements performed on day 21, the day of LA start, and again on the day of gonadotrophin start. The mean age was 30.6 +/- 3.9 years (range 23-37). Basal FSH was 5.4 +/- 1.9 IU/l (range 1.2-10.2). The mean preLA ovarian volume was 7.0 +/- 3.6 cm3 (left ovary 6.8 +/- 3.9, right ovary 7.1 +/- 3.8), compared to 6.3 +/- 4.2 cm3 postLA (left ovary 6.0 +/- 4.9, right ovary 6.5 +/- 4.8) (not significant). The mean number of small antral follicles noted in both ovaries was also unchanged after pituitary desensitization. Pituitary desensitization using LA had no effect on overall ovarian volume measurements. The total number of retrieved oocytes decreased with increasing age and decreasing ovarian volume.

Adult↗
PubMed1999

A safe, alternative technique for inferior turbinate reduction.

OBJECTIVE: Submucous resection of the inferior turbinates is a conventional technique for reducing their size to achieve patent nasal airways in situations where an enlarged turbinate contributes to airway obstruction. Many techniques and complications have been described in the past. We describe a new inferior turbinate reduction technique performed with powered instrumentation and assess its success and complication rates. STUDY DESIGN: A prospective study of 120 consecutive patients who underwent submucous resection of the inferior turbinates with a microdebrider. METHODS: Patient questionnaires were used for subjective assessment of symptoms before and after the procedure. We graded each patient's inferior turbinates for size from I to III before and 6 weeks after surgery for objective analysis. RESULTS: The common complications of standard submucous resection of inferior turbinates include excessive resection, postoperative bleeding, and crusting. The advantage of the microdebrider technique is the precise control of the amount of tissue and location of tissue that is removed on a submucosal plane. The complications encountered with this technique are limited to postoperative bleeding that occurred in 1.6% of patients. There was no crusting or excessive removal of tissue. CONCLUSION: The results show that submucous resection of inferior turbinates with a microdebrider is a safe method of achieving turbinate size reduction with minimal morbidity.

Adult↗
PubMed1999

Effects of dehydroepiandrosterone vs androstenedione supplementation in men.

PURPOSE: The purpose of this study was to compare the effects of short-term (12 wk) supplementation with androstenedione versus dehydroepiandrosterone (DHEA) on body composition, strength, and related hormones in middle-aged men. METHODS: A randomized, placebo-controlled, double-blind design was used to study 40 healthy, trained (>1 yr weight training) male subjects (mean +/- SD: age 48.1 +/- 3.9 yr; weight 79.8 +/- 9.8 kg). Subjects were randomly assigned to one of three groups: placebo (P), DHEA (D), or androstenedione (A). Supplements (50 mg capsules) were ingested two times daily for 12 wk. All testing, including venous blood samples, body composition, and performance, was conducted at three time points: presupplementation (1 d), at 6 wk, and postsupplementation (12 wk). RESULTS: Despite a small increase in lean body mass (0.8 +/- 0.4 and 0.5 +/- 0.3 kg) and mean strength (6.8 +/- 2.7 and 5.7 +/- 2.4 kg) in both D and A groups respectively, these changes were not significantly different from P. In D, there was a significantly greater increase in DHEA-S levels than in P (P < 0.05). There were no adverse side effects demonstrated during D or A supplementation including significant changes in PSA, liver function, or lipid levels (P < 0.05). CONCLUSIONS: The results of this study suggest that supplementation with 100 mg x d(-1) of either androstenedione or DHEA does not independently elicit a statistically significant increase in lean body mass, strength, or testosterone levels in healthy adult men over a 12-wk period.

Adult↗
PubMed1999

Oligospermic infertility associated with an androgen receptor mutation that disrupts interdomain and coactivator (TIF2) interactions.

Structural changes in the androgen receptor (AR) are one of the causes of defective spermatogenesis. We screened the AR gene of 173 infertile men with impaired spermatogenesis and identified 3 of them, unrelated, who each had a single adenine-->guanine transition that changed codon 886 in exon 8 from methionine to valine. This mutation was significantly associated with the severely oligospermic phenotype and was not detected in 400 control AR alleles. Despite the location of this substitution in the ligand-binding domain (LBD) of the AR, neither the genital skin fibroblasts of the subjects nor transfected cell types expressing the mutant receptor had any androgen-binding abnormality. However, the mutant receptor had a consistently (approximately 50%) reduced capacity to transactivate each of 2 different androgen-inducible reporter genes in 3 different cell lines. Deficient transactivation correlated with reduced binding of mutant AR complexes to androgen response elements. Coexpression of AR domain fragments in mammalian and yeast two-hybrid studies suggests that the mutation disrupts interactions of the LBD with another LBD, with the NH2-terminal transactivation domain, and with the transcriptional intermediary factor TIF2. These data suggest that a functional element centered around M886 has a role, not for ligand binding, but for interdomain and coactivator interactions culminating in the formation of a normal transcription complex.

Adult↗
PubMed1999

Decreased prevalence of retinopathy of prematurity in an inner-city hospital.

BACKGROUND AND OBJECTIVE: Improvements in care of low birth weight preterm infants has led to an increased survival of this population. It is not known whether the improved care has decreased the prevalence of retinopathy of prematurity (ROP), or whether the survival of even smaller infants has increased the prevalence. The purpose of this study was to determine the prevalence of ROP in an inner-city hospital during two time periods to determine whether the prevalence is changing. PATIENTS AND METHODS: The frequency and characteristics of ROP were compared between a 10-month observation period in 1995-1996 and published data from a similar time period in 1988-1989. RESULTS: There were fewer cases of ROP per live births less than 1500 g during 1995-1996 compared with 1988-1989 (P< .0001). This included both mild and severe ROP. When the two racial subgroups were examined separately, there was a statistically significant decline for the black (P < .001) but not the Hispanic population (P = .12). The Mantel-Haenszel chi2 test confirmed that the decline was statistically significant when the two groups were combined (P < .001). CONCLUSIONS: In this population, the prevalence of ROP appears to have decreased between 1995-1996 and 1988-1989. The subgroup experiencing the biggest decline was black infants. Other studies should be performed to investigate whether the frequency of ROP has also declined in other settings.

California↗
PubMed1998

Urea inhibits the Na-K pump in human erythrocytes.

Recent studies have established that urea alters the activity of several volume-sensitive cation transport pathways. However, it has remained unclear whether urea has any effect on transport pathways that are not volume-sensitive. We examined the effect of urea on Na-K pump in the human erythrocytes. In cells from nine subjects, 500 mM urea inhibited 52 +/- 10% of the pump activity measured as the ouabain-sensitive (OS) K influx. Urea inhibited the OS K influx reversibly, in a concentration-dependent manner. [3H] oubain binding, a measure of the number of Na-K pump sites remained unchanged with urea. Urea decreased the Vmax for ouabain-sensitive K influx, but did not alter the apparent K(m) for external K. Furthermore, urea did not alter the apparent K(m) for intracellular Na. The ion turnover per pump site was decreased in the presence of urea. Thus, physiologically relevant urea concentration inhibit the Na-K pump in human erythrocyte. The inhibition of the Na-K pump by urea suggests that the effects of urea may not be limited to volume-sensitive transporters, but may be more widespread.

Biological Transport↗
PubMed1998

NRP/B, a novel nuclear matrix protein, associates with p110(RB) and is involved in neuronal differentiation.

The nuclear matrix is defined as the insoluble framework of the nucleus and has been implicated in the regulation of gene expression, the cell cycle, and nuclear structural integrity via linkage to intermediate filaments of the cytoskeleton. We have discovered a novel nuclear matrix protein, NRP/B (nuclear restricted protein/brain), which contains two major structural elements: a BTB domain-like structure in the predicted NH2 terminus, and a "kelch motif" in the predicted COOH-terminal domain. NRP/B mRNA (5.5 kb) is predominantly expressed in human fetal and adult brain with minor expression in kidney and pancreas. During mouse embryogenesis, NRP/B mRNA expression is upregulated in the nervous system. The NRP/B protein is expressed in rat primary hippocampal neurons, but not in primary astrocytes. NRP/B expression was upregulated during the differentiation of murine Neuro 2A and human SH-SY5Y neuroblastoma cells. Overexpression of NRP/B in these cells augmented neuronal process formation. Treatment with antisense NRP/B oligodeoxynucleotides inhibited the neurite development of rat primary hippocampal neurons as well as the neuronal process formation during neuronal differentiation of PC-12 cells. Since the hypophosphorylated form of retinoblastoma protein (p110(RB)) is found to be associated with the nuclear matrix and overexpression of p110(RB) induces neuronal differentiation, we investigated whether NRP/B is associated with p110(RB). Both in vivo and in vitro experiments demonstrate that NRP/B can be phosphorylated and can bind to the functionally active hypophosphorylated form of the p110(RB) during neuronal differentiation of SH-SY5Y neuroblastoma cells induced by retinoic acid. Our studies indicate that NRP/B is a novel nuclear matrix protein, specifically expressed in primary neurons, that interacts with p110(RB) and participates in the regulation of neuronal process formation.

Adult↗
PubMed1998

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Anesthetics, Local↗