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Biomedical subjects

C Lee

Publications and source records attributed to C Lee.

At least 379 records · Page 21Linked to original sources

Expression of TGF-beta-1 mRNA and ultrastructural alterations in pharmacologically induced prolonged penile erection in a canine model.

PURPOSE: Transforming growth factor beta (TGF-beta) is known to induce fibrosis. Our objective was to study the role of TGF-beta as a possible mediator of fibrosis that may follow prolonged penile erection. MATERIALS AND METHODS: Prolonged penile erection was induced in seven adult male mongrel dogs by intracavernosal injection of papaverine into one of the corpora cavernosa while the other was used as a control. Intracavernosal pressure measurements were carried out prior to administration of papaverine and at the end of the procedure. Penile tissue was collected from anesthetized animals prior to euthanasia for histological and electron microscopic (EM) studies. RT-PCR was carried out for detection of mRNA on same tissue samples. RESULTS: The light microscopy showed stasis of blood in the cavernosal sinusoids. EM studies revealed sporadic endothelial defects, loss of plasma membrane integrity and cytoplasmic condensation. There was expression of TGF-beta1 mRNA in 66.7% of the experimental group compared with 16.7% of the control group. CONCLUSIONS: Pharmacologically induced low flow prolonged penile erection in canine models is associated with histomorphological changes in relatively short periods of time, suggesting that early therapeutic intervention is desirable. The gene expression for TGF-beta1 may be a mediator of fibrosis; therefore the use of anti-TGF-beta agents presents a possible tool for therapeutic intervention.

Animals↗

The pathogenesis of benign prostatic hyperplasia: a proposed hypothesis and critical evaluation.

PURPOSE: We used expanding observations regarding effects of testicular epididymal plasma and nonandrogenic testis factor(s) (NATF) on prostate growth to propose and evaluate a hypothesis regarding the development of benign prostatic hyperplasia (BPH) in man. MATERIALS AND METHODS: Current experimental data regarding the presence of NATF were reviewed. The potential for their exposure to the prostate by various routes was assessed. These observations were coupled with recognized anatomical, histological and epidemiological characteristics of BPH to construct a hypothesis regarding its pathogenesis. RESULTS: In vivo observations in man, rats and dogs supported the systemic secretion of NATF. These factors probably are, at least in part, spermatogenesis related. In vitro evaluation of the effect of spermatocele derived testicular epididymal plasma on human prostate stromal cells indicated the presence of androgen independent and androgen synergistic stromal growth promoters. These factors have potential local and systemic access to the prostate. The almost ubiquitous development of a regional, histologically variegated nodular growth occurring in the prostate in the androgen diminished environment of the aging man is compatible with local as well as systemic exposure to an age associated secretion of NATF. CONCLUSIONS: We propose that human BPH is an induced phenomenon that is usually initiated by local episodic exposure of periurethral prostate to mitogens secreted by the testis/epididymis. Once initiated, isolated or complex interacting proliferative stimuli from the testis/epididymis and a variety of other sources may achieve exposure to the prostate by several routes and simulate prostate growth.

Animals↗

A cadaveric and clinical evaluation of endoscopically assisted zygomatic fracture repair.

An endoscopic method of malar arch repair without a bicoronal incision has been recently described. To determine the effectiveness of this new technique, a cadaver study was performed to evaluate the capacity of this technique to (1) restore the anatomic position of a fractured malar arch, (2) rigidly fixate the arch, and (3) avoid injury to the frontal branch of the facial nerve. The technique of endoscopically assisted fracture repair was then applied to a clinical series of consecutive patients presenting with displaced zygomatic fractures with comminution at the malar arch. All cadaveric specimens repaired with this endoscopic technique demonstrated anatomic reduction and rigid fixation of the arch without disruption of the frontal branch of the facial nerve. In all clinical cases, four-point rigid plate fixation (zygomaticofrontal, infraorbital, malar arch, and zygomaticomaxillary buttress) was achieved endoscopically with limited access incisions. All clinical cases demonstrated excellent skeletal restoration of the zygoma on postoperative computed tomography scans. On clinical examination, facial symmetry and normal facial nerve function were observed in all patients after operation.

Cadaver↗

Endoscopic subcondylar fracture repair: functional, aesthetic, and radiographic outcomes.

An endoscopic method of mandibular subcondylar fracture repair has been described recently. To determine the effectiveness of this new technique, we longitudinally studied functional, aesthetic, and radiographic parameters following endoscopic repair of 22 subcondylar fractures in 20 patients. Restoration of mandibular function was achieved without postoperative maxillomandibular fixation. Premorbid occlusion was restored. Clinical jaw motion was found to progressively increase with a mean interincisal jaw opening of 43 mm achieved after the eighth postoperative week. Patients were pleased with the aesthetic restoration of their chin projection,jaw line, and the symmetric midline movement of the chin point onjaw opening. Anatomic fracture reduction with rigid plate fixation was confirmed on early postsurgical radiographs. Late radiographs showed fracture union without remodeling of the condylar head. Endoscopic subcondylar fracture repair was efficacious at functional, aesthetic, and radiographic restoration of the mandible.

Adult↗

Differential regulation of enolase during anaerobiosis in maize.

It was reported previously that enolase enzyme activity and ENO1 transcript levels are induced by anaerobic stress in maize (Zea mays). Here we show that not all isoforms of maize enolase are anaerobically induced. We cloned and sequenced a second enolase cDNA clone (pENO2) from maize. Sequence analysis showed that pENO2 shares 75.6% nucleotide and 89.5% deduced amino acid sequence identity with pENO1 and is encoded by a distinct gene. Expression of ENO2 is constitutive under aerobic conditions, whereas ENO1 levels are induced 10-fold in maize roots after 24 h of anaerobic treatment. Western-blot analysis and N-terminal sequencing of in vivo-labeled maize roots identified two major proteins selectively synthesized upon anaerobic stress as isozymes of enolase. We describe the expression of enolase in maize roots under anaerobic stress.

Amino Acid Sequence↗

MP1 encodes an abundant and highly antigenic cell wall mannoprotein in the pathogenic fungus Penicillium marneffei.

We cloned the MP1 gene, which encodes an abundant antigenic cell wall mannoprotein from the dimorphic pathogenic fungus Penicillium marneffei. MP1 is a unique gene without homologs in sequence databases. It codes for a protein, Mp1p, of 462 amino acid residues, with a few sequence features that are present in several cell wall proteins of Saccharomyces cerevisiae and Candida albicans. It contains two putative N glycosylation sites, a serine- and threonine-rich region for O glycosylation, a signal peptide, and a putative glycosylphosphatidylinositol attachment signal sequence. Specific anti-Mp1p antibody was generated with recombinant Mp1p protein purified from Escherichia coli to allow further characterization of Mp1p. Western blot analysis with anti-Mp1p antibody revealed that Mp1p has predominant bands with molecular masses of 58 and 90 kDa and that it belongs to a group of cell wall proteins that can be readily removed from yeast cell surfaces by glucanase digestion. In addition, Mp1p is an abundant yeast glycoprotein and has high affinity for concanavalin A, a characteristic indicative of a mannoprotein. Furthermore, ultrastructural analysis with immunogold staining indicated that Mp1p is present in the cell walls of the yeast, hyphae, and conidia of P. marneffei. Finally, it was observed that infected patients develop a specific antibody response against Mp1p, suggesting that this protein represents a good cell surface target for host humoral immunity.

Amino Acid Sequence↗

Detection of specific antibodies to an antigenic mannoprotein for diagnosis of Penicillium marneffei penicilliosis.

The disseminated and progressive fungal disease Penicillium marneffei penicilliosis is one of the most common infectious diseases in AIDS patients in Southeast Asia. To diagnose systemic penicilliosis, we developed an enzyme-linked immunosorbent assay (ELISA)-based antibody test with Mp1p, a purified recombinant antigenic mannoprotein of P. marneffei. Evaluation of the test with guinea pig sera against P. marneffei and other pathogenic fungi indicated that this assay was specific for P. marneffei. Clinical evaluation revealed that high levels of specific antibody were detected in two immunocompetent penicilliosis patients. Furthermore, approximately 80% (14 of 17) of the documented penicilliosis patients with human immunodeficiency virus tested positive for the specific antibody. No false-positive results were found for serum samples from 90 healthy blood donors, 20 patients with typhoid fever, and 55 patients with tuberculosis, indicating a high specificity of the test. Thus, this ELISA-based test for the detection of anti-Mp1p antibody can be of significant value as a diagnostic for penicilliosis.

AIDS-Related Opportunistic Infections↗

Circadian regulation of a Drosophila homolog of the mammalian Clock gene: PER and TIM function as positive regulators.

The Clock gene plays an essential role in the manifestation of circadian rhythms (approximately 24 h) in mice and is a member of the basic helix-loop-helix (bHLH) PER-ARNT-SIM (PAS) superfamily of transcription factors. Here we report the characterization of a novel Drosophila bHLH-PAS protein that is highly homologous to mammalian CLOCK. (Similar findings were recently described by Allada et al. Cell 93:791-804, 1998, and Darlington et al., Science 280:1599-1603, 1998.) Transcripts from this putative Clock ortholog (designated dClock) undergo daily rhythms in abundance that are antiphase to the cycling observed for the RNA products from the Drosophila melanogaster circadian clock genes period (per) and timeless (tim). Furthermore, dClock RNA cycling is abolished and the levels are at trough values in the absence of either PER or TIM, suggesting that these two proteins can function as transcriptional activators, a possibility which is in stark contrast to their previously characterized role in transcriptional autoinhibition. Finally, the temporal regulation of dClock expression is quickly perturbed by shifts in light-dark cycles, indicating that this molecular rhythm is closely connected to the photic entrainment pathway. The isolation of a Drosophila homolog of Clock together with the recent discovery of mammalian homologs of per indicate that there is high structural conservation in the integral components underlying circadian oscillators in Drosophila and mammals. Nevertheless, because mammalian Clock mRNA is constitutively expressed, our findings are a further example of striking differences in the regulation of putative circadian clock orthologs in different species.

Amino Acid Sequence↗

Prevalence of lower urinary tract symptoms in Korean men in a community-based study.

OBJECTIVE: The prevalence of lower urinary tract symptoms was determined in Korean men aged 50 and over. METHODS: A community-based, epidemiologic study was performed in Yonchon County, Korea. The Korean version of the International Prostate Symptom Score (I-PSS) was used to assess the severity of urinary symptoms in a representative sample of 514 men. RESULTS: Nocturia and weak stream were the most prevalent symptoms and urgency was the least. Overall, 23.2% of the men were moderately to severely symptomatic: 17.7% in the age group of 50-59 years, 23.3% in that of 60-69 years and 35.3% in that of 70 years and over. The proportion of severely symptomatic men approximately doubled with each decade of age. The 'quality of life' score showed a high correlation with the I-PSS. Our estimation indicated that in 1995 approximately 800,000 Korean men had moderate to severe lower urinary tract symptoms that were likely to be associated with benign prostatic hyperplasia. CONCLUSIONS: The prevalence of moderate to severe lower urinary tract symptoms in Korean men is substantially similar to that in Caucasians.

Aged↗

A proliferative effect of transforming growth factor-beta1 on a human prostate cancer cell line, TSU-Pr1.

Transforming growth factor -beta (TGF-beta) is growth inhibitory to many malignant cells, including prostate cancer cells. The present study reports an unusual observation in that TGF-beta is growth stimulatory to a human prostate cancer cell line, TSU-Pr1. The TSU-Pr1 line is highly aggressive and exhibits a rapid rate of proliferation in culture. These cells underwent further proliferation in response to TGF-beta1. Both type I and II receptors to TGF-beta (TPR-I, TPR-II) are expressed in TSU-Pr1 cells. Activation of a luciferase reporter gene, which contains a TGF-beta response element, confirmed that the TGF-beta receptors in TSU-Pr1 cells were functional. RT-PCR analysis and an ELISA assay determined that TSU-Pr1 cells secreted TGF-beta. In conclusion, TSU-Pr1 cells contain functional TGF-beta receptors but instead of the usual growth inhibition by TGF-beta1, these cells undergo proliferation. The present observation provides a proliferative role of TGF-beta in TSU-Pr1 cells, which may play a part in the aggressive phenotype of these cells and, perhaps other prostate cancer cells.

Cell Division↗

Immunohistochemical study of protein kinase C in the testes of cattle and pigs.

We investigated the expression of novel protein kinase C (PKC) delta and theta in the testes of pigs and cattle using Western blot and immunohistochemical analysis. PKC delta and theta are recognized in the testes of pigs and cattle by Western blot analysis. We found in immunohistochemical study that PCK delta was localized in the spermatids of seminiferous tubules, but not in the interstitial cells, while PKC theta was recognized only in the interstitial cells of the testes of in both species. These findings suggest that PKC delta and theta play an important role in the development of spermatozoa and the regulation of androgen in the testicular interstitial cells (probably Leydig cells), respectively.

Animals↗

Women's Health Australia: recruitment for a national longitudinal cohort study.

The Women's Health Australia (WHA) project is a longitudinal study of several cohorts of Australian women, which aims to examine the relationships between biological, psychological, social and lifestyle factors and women's physical health, emotional well-being, and their use of and satisfaction with health care. Using the Medicare database as a sampling frame (with oversampling of women from rural and remote areas), 106,000 women in the three age groups 18-23, 45-50 and 70-75 were sent an invitation to participate and a 24-page self-complete questionnaire. Reminder letters, a nation-wide publicity campaign, information brochures, a freecall number for inquiries, and the option of completing the questionnaire by telephone in English or in the respondent's own language, were used to encourage participation. Statutory regulations precluded telephone follow-up of non-respondents. Response rates were 41% (N = 14,792), 54% (N = 14,200) and 36% (N = 12,614) for the three age groups. Comparison with Australian census data indicated that the samples are reasonably representative of Australian women in these age groups, except for a somewhat higher representation of women who are married or in a defacto relationship, and of women with post-school education. The most common reason for non-participation was lack of interest or time. Personal circumstances, objections to the questionnaire or specific items in it, and concerns about confidentiality were the other main reasons. Recruitment of three representative age-group cohorts of women, and the maintenance of these cohorts over a number of years, will provide a valuable opportunity to examine associations over time between aspects of women's lives and their physical and emotional health and well-being.

Adult↗

Keratinocyte growth factor in the rat ventral prostate: androgen-independent expression.

Keratinocyte growth factor (KGF/FGF-7) is a stromally derived factor which exerts proliferative and differentiating effects on a variety of epithelial cells. Results of recent studies utilizing in vitro methods such as tissue culture and organ culture have suggested that KGF may act as a paracrine mediator of androgen-induced growth and development of the prostate and seminal vesicle. We undertook the present study to determine the distribution of KGF in relation to the functional regions of the rat prostatic ductal system, and whether KGF expression is influenced by androgen in vivo. Immunohistochemical staining revealed KGF to be present in the stroma throughout the prostate, regardless of the functional region, and staining for KGF remained high through 21 days post-castration. Message for KGF could also be detected by reverse transcriptase-PCR analysis of prostate stromal cells isolated from 4- and 21-day castrated animals, and no gross change in message level was observed following castration. Furthermore, no significant change in either stromal staining or message for KGF was observed in newborn rat prostates 10 days after castration, suggesting a similar regulatory mechanism for KGF in the adult and immature prostate. Epithelial staining for KGF decreased following castration, and greatly increased upon androgen replacement, possibly indicating a change in KGF internalization. These observations suggest that the presence of KGF protein is not related to functional differences in the prostate epithelium, and that expression of KGF in vivo is not greatly influenced by androgen.

Androgens↗

Multiple sites of loss of heterozygosity on chromosome 8 in human breast cancer has differential correlation with clinical parameters.

Recent studies have shown that chromosome 8p21-22 is the main site of frequent loss of heterozygosity (LOH) in breast cancers. However, the detailed molecular analysis of chromosome 8 so far in breast cancer has been variable. Most of the literature pertaining to LOH in breast cancer is mainly on short arm of chromosome 8. In the present study, we have examined LOH on both short and long arm of chromosome 8 using fifteen different polymorphic DNA markers in microdissected samples of normal breast epithelium and carcinoma from the same patients. For this purpose, DNA was extracted from the microdissected normal and tumor cells of 66 breast cancers, amplified by PCR and analyzed for LOH on chromosome 8 using fifteen different polymorphic DNA markers (D8S264, D8S298, D8S535, D8S255, D8S1098, D8S589, D8S567, D8S591, D8S285, D8S1102, D8S1763, D8S260, D8S530, D8S1772, and D8S1844). Expression of estrogen receptor, progesterone receptors, and p53 antigens was determined by immunohistochemistry using specific monoclonal antibodies. The results of this study suggest that LOH on chromosome 8 was identified in 40 of 66 cases (61%) with at least one marker. Three distinct regions of loss detected were: i) at 8p12, at loci between D8S535 and D8S255; ii) at 8p11, on loci D8S567, D8S591, D8S285, and D8S1102; iii) at 8q11-12, on loci D8S1763, D8S260 and D8S530. We found 45% (30 out of 66 informative cases) of the tumors showed LOH at 8p12; 52% (34 out of 66 informative cases) had LOH at 8p11; and 39% (26 out of 66 informative cases) had LOH at 8q11-12. Deletion at 8q11-12 was significantly correlated with the grade of the breast cancer specimens. Moderate to poorly differentiated specimens had higher incidence of LOH at 8q11-12 as compared to well differentiated specimens. Deletion at 8p12 and 8p11 was significantly higher in clinical stages III and IV of breast cancer tissues as compared to stage I and II cases. Tissues with lymph node involvement showed higher incidence of LOH at 8p12 as compared to the tissues with no lymph node involvement. There was no correlation of LOH at these loci with either the age of the patients, tumor size, BrdU labeling index, expression of estrogen receptor, progesterone receptor, and p53 in breast cancer specimens. These experiments, for the first time, report multiple sites of LOH on chromosome 8 in human breast cancer, and these deletions have differential correlation with clinical parameters of breast cancer samples.

Breast Neoplasms↗

Microsatellite instability in an animal model of mammary carcinogenesis.

Alterations in the length of simple repetitive genomic sequences (microsatellite instability, MSI) may characterize a distinct mechanism of mammary carcinogenesis. In order to investigate whether MSI is associated with chemically-induced mammary carcinogenesis in the rat, 30 microdissected mammary carcinomas were analyzed using 27 different microsatellite markers from chromosomes 1, 3, 5, 7 and 8. DNA was extracted from rat mammary cancer and adjoining microscopically normal tissues from the same slide, amplified by PCR, using different polymorphic DNA markers and the reaction products were analyzed for microsatellite instability. The results of this study indicate that 30% of cases (9 out of 30) showed microsatellite instability at a minimum of 1 locus. Three cases (out of 30) showed microsatellite instability at only three loci or less, called MSI-L (low frequency MSI). Six cases (out of 30) showed MSI at four loci or more, called MSI-H (high frequency MSI). Six cases showed MSI at D5Mit11 and D5Mgh3 loci, five cases showed MSI at D1Mit14, D1Mgh6, D5Mgh5 and D8Mgh10 loci, four cases had MSI at D1Mgh2, and D3Mgh7 loci, three cases had MSI at D3Mit3, D3Mgh5, D7Mgh1 loci, two cases had MSI at D7Mit11 locus and one case had MSI at D3Mgh9 locus. The results of these experiments suggest that MSI may be an important etiological event in the pathophysiology of mammary carcinogenesis.

Animals↗