BACKGROUND: Eosinophil cationic protein (ECP), one of the eosinophil granule proteins released during allergic reactions, may play a major role in the allergic inflammatory process. The measurement of ECP in serum may be a useful indicator of eosinophil activity in ongoing inflammatory processes. We investigated the clinical utility of ECP measurement in serum in patients with bronchial asthma, allergic rhinitis and atopic dermatitis, after standardizing sample processing. METHODS: We measured the serum ECP levels in patients with bronchial asthma (n=38), chronic obstructive pulmonary diseases (COPD) (n=13), respiratory symptoms (n=19), allergic rhinitis (n=26), non-allergic rhinitis (n=24), and atopic dermatitis (n=10) and in normal healthy controls (n=16) by the fluoroenzyme immunoassay using Pharmacia CAP System, and evaluated the correlation between ECP level and blood eosinophil number, or ECP and IgE levels. Blood eosinophil number was counted by the automated cell counter. RESULTS: Serum ECP levels were significantly higher in patients with bronchial asthma (15.6+/- 12.6 g/L), COPD (13.3+/-7.2 g/L), allergic rhinitis (23.8+/-13.2 g/L), and atopic dermatitis (20.6+/- 18.4 g/L) than in normal controls (7.5+/-4.2 g/L) (P <0.05). ECP levels were also significantly higher in patients with bronchial asthma and COPD than in patients with simple respiratory symptoms (6.9+/-4.7 g/L), whose ECP levels did not statistically differ from those in normal controls. ECP levels were also significantly higher in patients with allergic rhinitis than in patients with non-allergic rhinitis (9.5+/-5.1 g/L), whose ECP levels did not statistically differ from those in normal controls. Serum ECP level and eosinophil number in peripheral blood were correlated only in patients with bronchial asthma (r=0.53, P <0.01) and no correlation between ECP and IgE levels was found in all of the patients. CONCLUSIONS: ECP is the one of the secretory components released from the eosinophil granule and measurement of ECP in serum might be one of the noninvasive tool to assess the activity in relation to eosinophil involvement in various allergic diseases.
Asthma ; Cell Count ; Dermatitis, Atopic ; Eosinophil Cationic Protein* ; Eosinophil Granule Proteins ; Eosinophils ; Humans ; Hypersensitivity ; Immunoassay ; Immunoglobulin E ; Lung Diseases, Obstructive ; Pulmonary Disease, Chronic Obstructive ; Rhinitis

OBJECTIVE: Identification of the regulatory mechanism for arrest and initiation of primordial follicular growth is crucial for female fertility. Previously, we found 15 expressed sequence tags (ESTs) that were specifically abundant in the day-5-subtracted cDNA library and that the B357 clone was novel. The present study was conducted to obtain the whole sequence of the novel gene including B357 and to characterize its mRNA and protein expression in mouse ovary and testis. METHODS: The extended sequence of the 2,965-bp cDNA fragment for the clone B357 was named 5-day-ovary-specific gene-1 (5DOS1) and submitted to GenBank (accession number AY751521). Expression of 5DOS1 was characterized in both female and male gonads at various developmental stages by Northern blotting, real-time RT-PCR, in situ hybridization, Western blotting, and immunohistochemistry. RESULTS: The 5DOS1 transcript was highly expressed in the adult testis, brain, and muscle as compared to the other tissues. In the ovary, the 5DOS1 transcript was detected in all oocytes from primordial to antral follicles, and highly expressed at day 5 after birth and decreased thereafter. In contrast, expression of 5DOS1 showed a gradual increase during testicular development and its expression was limited to various stages of male germ cells except spermatogonia. CONCLUSIONS: This is the first report on the expression and characterization of the 5DOS1 gene in the mouse gonads. Further functional analysis of the 5DOS1 protein will be required to predict its role in gametogenesis.
Adult ; Animals ; Blotting, Northern ; Blotting, Western ; Brain ; Clone Cells ; Cluster Analysis* ; Databases, Nucleic Acid ; DNA, Complementary ; Expressed Sequence Tags ; Female ; Fertility ; Gametogenesis ; Gene Library ; Germ Cells ; Gonads ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Male ; Mice* ; Oocytes ; Ovary* ; Parturition ; RNA, Messenger ; Spermatogonia ; Testis*

BACKGROUND: The product of oxygen-free radicals inf1ict oxidative injuries on healthy cells. Antioxidants such as superoxide dismutase(SOD), glutathione peroxidase, and reduced glutathione(GSH) are present in almost all cells and play important roles in metabolism, transport, and cellular protection. We measured blood GSH levels in healthy controls and patients with non insulin dependent diabetes mellitus(NIDDM) for evaluation of the clinical usefulness of GSH. METHODS: Erythrocyte GSH levels were measured in fifty healthy controls and thirty NIDDM patients with diabetic retinopathies by Beutler's method. We also tested within-run precision, between-run precision, linearity and recovery rate to evaluate this method measuring erythrocyte GSH levels. RESULTS: The GSH levels (mean +/-SD) of NIDDM patients (5.03+/-0.67mumo1/Hb) were significantly lower than those of healthy control group (6.46+/-0.85mumo1/Hb)(P<0.001). The results of within-run precision and between-run precision when stored at 4degrees Cwere excellent (coefficient of variation were 2.79% and 2.42%, respectively), however, when stored at the room temperature the GSH levels were sharply declined. The linearity and recovery rate were acceptable. CONCLUSIONS: The prescision, linearity, and recovery rate of GSH measurement were excellent. The GSH levels in NIDDM patient group were reduced, and this probably contributes to the defective defense mechanism against increased oxidative stress. Additional measurement of other antioxidants such as superoxide dismutase and glutathione Peroxidase may be required to clarify the pathologic significance of glutathione metabolism in various diseases.
Antioxidants ; Diabetes Mellitus, Type 2 ; Diabetic Retinopathy ; Erythrocytes* ; Glutathione Peroxidase ; Glutathione* ; Humans ; Insulin ; Metabolism ; Oxidative Stress ; Superoxide Dismutase ; Superoxides

OBJECTIVE: Recently, microdissection of tissue sections has been used increasingly for the isolation of morphologically identified homogeneous cell populations, thus overcoming the obstacle of tissue complexity for the analysis cell-specific expression of macromolecules. The aim of the present study was to establish the minimal conditions required for the RNA extraction and amplification from the cells captured by the laser captured microdissection. METHODS: Mouse ovaries were fixed and cut into serial sections (7 micrometer thickness). Oocytes were captured by laser captured microdissection (LCM) method by using PixCell IITM system. The frozen sections were fixed in 70% ethanol and stained with hematoxylin and eosin, while the paraffin sections were stained with Multiple stain. Sections were dehydrated in graded alcohols followed by xylene and air-dried for 20 min prior to LCM. All reactions were performed in ribonuclease free solutions to prevent RNA degradation. After LCM, total RNA extraction from the captured oocytes was performed using the guanidinium isothiocyanate (GITC) solution, and subsequently evaluated by reverse transcriptase -polymerase chain reaction (RT-PCR) for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). RESULTS: With the frozen sections, detection of the GAPDH mRNA expression in the number of captured 25 oocytes were not repeatable, but the expression was always detectable from 50 oocytes. With 25 oocytes, at least 27 PCR cycles were required, whereas with 50 oocytes, 21 cycles were enough to detect GAPDH expression. Amount of the primary cDNA required for RT-PCR was reduced down to at least 0.25 microl with 50 oocytes, thus the resting 19.75 microl cDNA can be used for the testing other interested gene expression. Tissue-to-slide, tissue-to-tissue forces were very high in the paraffin sections, thus the greater number of cell procurement was required than the frozen sections. CONCLUSION: We have described a method for analyzing gene expression at the RNA level with the homogeneously microdissected cells from the small amount of tissues with complexity. We found that LCM coupled with RT-PCR could detect housekeeping gene expression in 50 oocytes captured. This technique can be easily applied for the study of gene expression with the small amount of tissues.
Alcohols ; Animals ; DNA, Complementary ; Eosine Yellowish-(YS) ; Ethanol ; Female ; Frozen Sections ; Gene Expression* ; Genes, Essential ; Guanidine ; Hematoxylin ; Mice ; Microdissection* ; Oocytes* ; Ovary ; Paraffin ; Polymerase Chain Reaction ; Ribonucleases ; RNA Stability ; RNA* ; RNA, Messenger ; RNA-Directed DNA Polymerase ; Xylenes

PURPOSE: This study was intended to investigate the types and seriousness of the couvade syndrome, pregnancy-related physical and psychological symptoms among expectant fathers whose spouses were pregnant. METHOD: The subject was consists of 100 expectant fathers at one hospital in Seoul, Korea. The pregnant women had not been diagnosed any medical complication. Data were analyzed by SPSS/PC program. RESULT: 1) The total mean score was 1.85: the mean score of perceived physical symptoms (1.87) revealed higher than the mean score of psychological symptoms (1.81). 2) With the respect to the general characteristics of subjects, there were statistically significant correlations between subject's level of education and couvade symptoms (r=-.209, p=.037), gestational age and couvade symptoms (r=-.227, p=.023), family total income and couvade symptoms (r=-.198, p=.048), perceived self health status and couvade symptoms (r=-.254, p=.011). 3) With the respect to the general characteristics of subjects, there were statistically significant differences in pregnant woman's age (t=1.363, p=.044),occupation of subject (F=3.594, p= .009), educational level of subject (t=3.506, p=.002), family total income (F=16.822, p= .000), perceived self health status (F=3.151, p=.047). CONCLUSION: Couvade syndrome is an issue for nurses who perform an important role in the care of pregnant women and their spouses.
Education ; Fathers ; Female ; Gestational Age ; Humans ; Korea ; Pregnant Women ; Seoul ; Spouses*

BACKGROUND: Although normal vascular endothelium prevents adhesion and aggregation of platelets by the release of nitric oxide (NO) and prostacyclin, circulating blood cells, such as polymorphonuclear leukocytes (PMNs) and mononuclear leukocytes (ML) may be considered to be also important in modulating platelet aggregation. Recently, nitric oxide synthase (NOS) activity was found in PMNs and ML, so these cells can also release NO to inhibit platelet aggregation. We studied platelet-ML interactions using an experimental model in which isolated ML were placed in the aggregometer in contact with human platelets, stimulated by collagen. METHODS: Platelet count in platelet-rich plasma (PRP) was adjusted to approximately 300x109/L. ML were separated using Ficoll-Hypaque (specific gravity 1.077) and finally resuspended at 1, 3 and 5x109/L, in Tyrode albumin buffer (TAB), respectively. Platelet aggregation was measured with Chrono-Log Aggregometer (USA) after adding variable numbers of the ML, stimulating with 2.5, 5 and 10 microgram/mL of collagen. Mechanisms of ML to inhibit the platelet aggregation were evaluated after incubating the ML with 10 micrometer indomethacin and 300 micrometer NG-monomethyl-L-arginine (L-NMMA). RESULTS: Non-stimulated ML (3x109/L) inhibited (43.2 +/- 19.6 versus TAB control 69.2 +/- 10.7% transmission) the platelet aggregation induced by 2.5 microgram/mL of collagen. The inhibition was not attenuated by increasing the concentration of collagen from 5.0 microgram/ mL (50.1 +/- 18.0% versus TAB control 75.5 +/- 13.1%, P<0.001) to 10 microgram/mL (62.9 +/- 17.3% versus TAB contol 82.3 +/- 12.6%, P<0.01). In addition, it was dependent on the number of ML and incubation time. While preincubation of the ML with indomethcin did not affect the antiaggregating capacity of the ML (63.4 +/- 11.1 versus TAB control 73.3 +/- 7.3%), preincubation of the ML with L-NMMA slightly inhibit the antiaggregating capacity of the ML (86.6 +/- 6.8 versus TAB control 73.3 +/- 7.3%). CONCLUSION: These findings suggest that blood ML inhibited the collagen-induced platelet aggregation, of which mechanism appears to be only partly dependent on NO and to be independent on prostaglandins. Release of other substances affecting platelet aggregation from ML requires to be clarified. Using our experimental model, it has been demonstrated that cell-cell contact may facilitate the exchange of a wide array of mediators between platelets and ML which may influence the cellular responses. This experimental model thus allows to study interactions between platelets and ML.
Blood Cells ; Blood Platelets* ; Collagen ; Endothelium, Vascular ; Epoprostenol ; Gravitation ; Humans ; Indomethacin ; Leukocytes, Mononuclear* ; Models, Theoretical ; Neutrophils ; Nitric Oxide ; Nitric Oxide Synthase ; omega-N-Methylarginine ; Platelet Aggregation* ; Platelet Count ; Platelet-Rich Plasma ; Prostaglandins

Acinic cell carcinoma (ACC) of the breast is extremely rare and is characterized by widespread acinar cell-like differentiation. We report of a 39-year-old woman presented with a palpable breast mass with significant morphological, immunohistochemical and ultrastructural findings. Histologically, ACC showed a diffuse glandular infiltrative pattern, with small acinar or glandular structures mixed with solid nests. Neoplastic cells were monotonous proliferation of cells with a granular or clear cytoplasm, resembling acinar cells of the salivary glands or Paneth cells. Both glandular and solid tumor cell populations were strongly positive for lysozyme and alpha-1-antitrypsin.
Acinar Cells ; Adult ; Breast ; Breast Neoplasms ; Carcinoma, Acinar Cell ; Cytoplasm ; Female ; Humans ; Immunohistochemistry ; Microscopy, Electron ; Muramidase ; Paneth Cells ; Salivary Glands

PURPOSE: To analyze the incidence, clinical characteristics and postoperative prognosis of simultaneous bilateral rhegmatogenous retinal detachment (SRD). METHODS: We retrospectively analyzed the medical records of 22 patients who visited the Department of Ophthalmology, Yeungnam University College of Medicine, for treatment of SRD and who had been in regular surveillance for at least 6 months. RESULTS: The incidence of SRD was 22 patients among 792 (2.8%). Eleven were male and 11 were female, and the average age was 27.6 years. Eighteen patients (81.8%) complained of visual disturbance and visual field defect of one eye, and myopia of more than -4.00D was noted in 30 eyes (68.2%). The size of the detached area was 2 quadrants in 19 eyes (43.2%). The most common type of retinal break was atrophic hole with lattice degeneration. The most common location of the break was the inferotemporal quadrant. The anatomical success rate of primary operation was 91.9% (34 of 37 eyes). CONCLUSIONS: Simultaneous bilateral rhegmatogenous retinal detachment was associated with young age, and myopia. As for the retinal break, the most common type was atrophic hole with lattice degeneration, and the most common location was the inferotemporal quadrant.
Female ; Male ; Humans ; Incidence

OBJECTIVES: Previously, we sought to compile a list of genes expressed during early folliculogenesis by using cDNA microarray to investigate follicular gene expression and changes during primordialprimary follicle transition and development of secondary follicles (Yoon et al., 2005). Among those genes, a group of genes related to the cell size growth was characterized during the ovarian development in the present study. METHODS: We determined ovarian expression pattern of six genes related to the cell size growth (cyr61, emp1, fhl1, socs2, wig1 and wisp1) and extended into CCN family (connective tissue growth factor/cysteine-rich 61/nephroblastoma-overexpressed), ctgf, nov, wisp2, wisp3, including cyr61 and wisp1 genes. Expression of mRNA and protein according to the ovarian developmental stage was evaluated by in situ hybridization, and/or semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry, respectively. RESULTS: Among 6 genes related to the cell size growth, cyr61 and wisp1 mRNA was detected only in oocytes in the postnatal day5 mouse ovaries. cyr61 mRNA expression was limited to the nucleolus of oocytes, while wisp1 was expressed in the cytoplasm and nucleolus of oocytes, except nucleus. cyr61 mRNA expression, however, was found in granulosa cells from secondary follicles. The rest 4 genes in the cell size growth group were detected in oocytes, granulosa and theca cells. Cyr61 and Wisp1 proteins were expressed in the oocyte cytoplasm from primordial follicle stage. Especially, Cyr61 protein was detected in pre-granulosa cells, Wisp1 protein was not. By using RT-PCR, we evaluated and decided that Cyr61 protein is produced by their own mRNA in pre-granulosa cells that was not detected by in situ hybridization. cyr61 and wisp1 genes are happen to be the CCN family members. The other members of CCN family were also studied, but their expression was detected in oocytes, granulose and theca cells. CONCLUSIONS: We firstly characterized the ovarian expression of genes related to the cell size growth and CCN family according to the early folliculogenesis. Cyr61 protein expression in the pre-granulosa cells is profound in meaning. Further functional analysis for cyr61 in early folliculogenesis is under investigation.
Animals ; Cell Enlargement* ; Cell Size* ; Cysteine-Rich Protein 61 ; Cytoplasm ; Female ; Gene Expression ; Genes, vif ; Granulosa Cells ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Mice* ; Oligonucleotide Array Sequence Analysis ; Oocytes ; Ovary ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger ; Theca Cells

PURPOSE: To evaluate the efficacy and long-term prognosis of pars plana vitrectomy and Ahmed valve implantation for intractable glaucoma comorbid with retinal disorders. METHODS: A retrospective review of 34 eyes of 30 patients receiving pars plana vitrectomy and Ahmed valve implantation for intractable glaucoma comorbid with retinal disorders was performed. Preoperative and postoperative intraocular pressure (IOP) and visual acuity, the usage of IOP-lowering medications and postoperative complications, and surgical success rate were evaluated. RESULTS: IOP and the number of IOP-lowering medications showed a significant decrease after pars plana vitrectomy and Ahmed valve implantation, as compared to before surgery (p < 0.001). The success rate was 88%, 88%, 84% and 85% postoperatively at 1 month, 6 months, 1 year and 3 years respectively. The mean follow up period was 24.21 +/- 14.99 months. Complications related to surgery included hyphema in 2 eyes, recurrent corneal epithelial erosion and defect in 2 eyes, corneal ulcer in 2 eyes and vitreous hemorrhage in 4 eyes. Visual acuity improved in 14 eyes (41.1%), no changes in 13 eyes (38.2%) and decreased in 7 eyes (20.6%). CONCLUSIONS: Pars plana vitrectomy and Ahmed valve implantation for intractable glaucoma comorbid with retinal disorders show long-term efficacy in lowering IOP.
Corneal Ulcer ; Eye ; Follow-Up Studies ; Glaucoma ; Humans ; Hyphema ; Intraocular Pressure ; Postoperative Complications ; Prognosis ; Retinaldehyde ; Retrospective Studies ; Visual Acuity ; Vitrectomy ; Vitreous Hemorrhage