Nuclear Localization of Fibroblast Growth Factor Receptor 1 in Breast Cancer Cells Interacting with Cancer Associated Fibroblasts

Cancer-associated fibroblasts (CAFs) represent a major component of the tumor microenvironment and interplay with cancer cells by secreting cytokines, growth factors and extracellular matrix proteins. When estrogen receptor-negative breast cancer MDAMB-231 cells were treated with the CAF-conditioned medium (CAF-CM), Akt and STAT3 involved in cell proliferation and survival were activated through phosphorylation. CAFs secrete fibroblast growth factor 2 (FGF2), thereby stimulating breast cancer cell progression. Akt activation induced by CAF-CM in MDA-MB-231 cells was abolished when FGF2-neutralizing antibody was added.Treatment of MDA-MB-231 cells directly with FGF2 enhanced the phosphorylation of Akt and the FGF receptor (FGFR) substrate, FRS2α. These events were abrogated by siRNA-mediated silencing of FGFR1. In a xenograft mouse model, co-injection of MDAMB-231 cells with activated fibroblasts expressing FGF2 dramatically enhanced activation of Akt. Stable knockdown of FGFR1 blunted Akt phosphorylation in xenograft tumors. MDA-MB-231 cells co-cultured with CAFs or directly stimulated with FGF2 exhibited enhanced nuclear localization of FGFR1. Notably, FGF2 stimulation produced reactive oxygen species (ROS) accumulation in MDA-MB-231 cells, and FGF2-induced nuclear accumulation of FGFR1 was abrogated by the ROS scavenging agent, N-acetylcysteine.

Genistein Inhibits Proliferation of BRCA1 Mutated Breast Cancer Cells: The GPR30-Akt Axis as a Potential Target

BACKGROUND: BRCA1 mutated breast cancer cells exhibit the elevated cell proliferation and the higher metastatic potential. G protein-coupled receptor 30 (GPR30) has been shown to regulate growth of hormonally responsive cancers, such as ovarian and breast cancers, and high expression of GPR30 is found in estrogen receptor (ER)-negative breast cancer cells. ER-negative breast cancer patients often have a mutation in the tumor suppressor gene, BRCA1. This study explored antiproliferative effects of genistein, a chemopreventive isoflavone present in legumes, and underlying molecular mechanisms in triple negative breast cancer cells with or without functionally active BRCA1.METHODS: Expression of BRCA1, GPR30 and Nrf2 was measured by Western blot analysis. Reactive oxygen species (ROS) accumulation was monitored by using the fluorescence-generating probe, 2’,7’-dichlorofluorescein diacetate. The effects of genistein on breast cancer cell viability and proliferation were assessed by the MTT, migration and clonogenic assays.RESULTS: The expression of GPR30 was dramatically elevated at both transcriptional and translational levels in BRCA1 mutated breast cancer cells compared to cells with wild-type BRCA1. Notably, there was diminished Akt phosporylation in GPR30 silenced cells. Treatment of BRCA1 silenced breast cancer cells with genistein resulted in the down-regulation of GPR30 expression and the inhibition of Akt phosphorylation as well as the reduced cell viability, migration and colony formation. Genistein caused cell cycle arrest at the G₂/M phase in BRCA1-mutant cells through down-regulation of cyclin B1 expression. Furthermore, BRCA1-mutant breast cancer cells exhibited higher levels of intracellular ROS than those in the wild-type cells. Genistein treatment lowered the ROS levels through up-regulation of Nrf2 expression.CONCLUSIONS: Lack of functional BRCA1 activates GPR30 signaling, thereby stimulating Akt phosphorylation and cell proliferation. Genistein induces G2/M phase arrest by down-regulating cyclin B1 expression, which is attributable to its suppression of GPR30 activation and Akt phosphorylation in BRCA1 impaired breast cancer cells.

Physiological Functionalities of Vitis hybrid (Sheridan)-Rubus coreanus Red Wine Made by Saccharomyces cerevisiae

Vitis hybrid (Sheridan)-Robus coreanus red wine was vinified by fermentation of a mixture of Vitis hybrid and Robus coreanus must at 25degrees C for 10 days. The Vitis hybrid-Robus coreanus red wine had ethanol contents of 10.9%. It had high antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity of 57.8% and antioxidant activity of 64.8%. Changes in the physicochemical properties and functionality of the Vitis hybrid-Robus coreanus red wine was investigated during a post-fermentation period of three months. The ACE inhibitory activity of the red wine increased as the post-fermentation period prolonged, and showed the highest ACE inhibitory activity of 70.4% 60 days post-fermentation. However, the antioxidant activity declined significantly to 47.2% during the post-fermentation period of 60 days. In terms of sensory evaluation, the Vitis hybrid-Robus coreanus red wine had the best acceptability 60 days post-fermentation.

Antihypertensive Angiotensin I-Converting Enzyme Inhibitory Activity and Antioxidant Activity of Vitis hybrid-Vitis coignetiae Red Wine Made with Saccharomyces cerevisiae

A Vitis hybrid-Vitis coignetiae red wine was vinified by fermentation of a mixture of a Vitis hybrid-Vitis coignetiae must with Saccharomyces cerevisiae KCTC 7904 at 25degrees C for 10 days. The Vitis hybrid-Vitis coignetiae red wine showed high antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity (67.8%) and antioxidant activity (76.7%). The antihypertensive ACE inhibitor in the Vitis hybrid-Vitis coignetiae red wine was partially purified by solid phase extraction chromatography, and its ACE inhibitory activity yielded an IC50 of 1.8 mg/mL. Six kinds of oligopeptides, including five new kinds, were contained in the partially purified ACE inhibitor fraction from the red wine after 10 days of fermentation. Antioxidant activity decreased significantly from 76.7% to 40.5% when the post-fermentation period was prolonged to 30 days.

Digestion Pattern of Antihypertensive Angiotensin I-Converting Enzyme Inhibitory Peptides from Saccharomyces cerevisiae in a Successive Simulated Gastricintestinal Bioreactor

A cell-free extract of Saccharomyces cerevisiae containing the angiotensin I-converting enzyme (ACE) inhibitory peptide was treated in a successive simulated gastric-intestinal bioreactor (step 1: amylase digestion, step 2: gastric fluid digestion, step 3: intestinal fluid digestion) to illustrate the absorption pattern of antihypertensive ACE inhibitory peptide, and the ACE inhibitory activities of each step were determined. Total ACE inhibitory activities of step 1, step 2, and step 3 were 55.96%, 80.09%, and 76.77%, respectively. The peptide sequence of each steps was analyzed by MS/MS spectrophotometry. Eleven kinds of representative peptide sequences were conserved in each step, and representative new peptides including RLPTESVPEPK were identified in step 3.

Extraction and Characteristics of Anti-obesity Lipase Inhibitor from Phellinus linteus

To develop a potent anti-obesity lipase inhibitor from mushroom, the lipase inhibitory activities of various mushroom extracts were determined. Methanol extracts from Phellinus linteus fruiting body exhibited the highest lipase inhibitory activity (72.8%). The inhibitor was maximally extracted by treatment of a P. linteus fruiting body with 80% methanol at 40degrees C for 24 hr. After partial purification by systematic solvent extraction, the inhibitor was stable in the range of 40~80degrees C and pH 2.0~9.0. In addition to lipase inhibitory activity, the inhibitor showed 59.4% of superoxide dismutase-like activity and 56.3% of acetylcholinesterase inhibitory activity.

Production of Ginsenoside-Rg3 from Lipomyces starkeyi Grown on Ginseng-Steaming Effluent

To produce ginsenoside-Rg3 enriched yeast from ginseng-steaming effluent (GSE), Lipomyces starkeyi, which tends to grow well in GSE, was cultured in sterilized GSE and its growth and production of ginsenoside-Rg3 were determined. Growth of L. starkeyi was 86.1 mg per g GSE and its ginsenoside-Rg3 contents was 0.013 mg per g GSE.

Production of Antihypertensive Angiotensin I-Converting Enzyme Inhibitor-Enriched Edible Yeast Using Gugija (Lycium chinesis Mill)

To produce bioactive compound enriched yeast using medicinal Gugiga (Lycium chinensis Mill), several edible Saccharomyces species were cultured in Gugija extracts added yeast extract, peptone and dextrose medium (GE - YEPD medium) at 30degrees C for 24 hr, and their growth were determined. Growth of Saccharomyces cerevisiae K-7 and Sacchromyces cerevisiae ACTC 7904 were better than those of the other yeasts. Two yeasts were selected and then determined their some physiological functionalities after cultivated the yeasts in the GE - YEPD medium and compared those grown on YEPD medium. Antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity of S. cerevisiae K-7 grown on GE - YEPD medium was about 20% higher than that grown on YEPD medium. Superoxide dismutase-like activity of S. cerevisiae ACTC 7904 was also about 12% more high. However, the other physiological functionalities were almost same or lower. Optimal addition concentration of Gugija extract was 10%, and maximally growth and ACE inhibitory activity of S. cerevisiae K-7 were shown when the strain was cultured in 10% Gugija extracts containing YEPD medium at 30degrees C for 12 hr.

Reflux Symptom Index(RSI) and Reflux Finding Score(RFS) of Persons Taking Health Checkup and Their Relationship with Gastrofiberscopic Findings / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Few studies were performed regarding laryngopharyngeal reflux symptoms and reflux findings in persons taking health checkup. The purpose of this study was to evaluate laryngopharyngeal reflux symptom index (RSI) and reflux finding score (RFS) at a healthcare center and their relationship with gastrofiberscopic findings. SUBJECTS AND METHOD: This study included 157 persons taking healthcare program which contained laryngoscopic examination between February 2006 and June 2006. The data of each patients were collected by questionnaire, fill-up sheets of laryngoscopic findings and a retrospective review of medical record. RESULTS: The male to female ratio was about 4 : 1. The mean age was 47 years. The mean values of RSI and RFS were 2.95 and 3.85. Seven (4.5%) persons had RSI more than 13, and 31 (19.7%) persons had RFS more than 7. On gastrofiberscopy, 23 (14.6%) persons had reflux esophagitis, and their RSI were significantly higher than those who did not have reflux esophagitis, and had frequent gastroesophageal reflux symptoms. In persons with abnormal RSI or RFS, only 29% had reflux esophagitis on gastrofiberscopy. Age was inversely related with RSI and smoking was positively related with RFS. CONCLUSION: We suggest that if a person was found to have abnormally elevated RSI or RFS, a diagnostic evaluation for LPR should be considered irrespective of the presence of reflux esophagitis on gastrofiberscopy. In a person with reflux esophagitis on gastrofiberscopy, he or she showed high RSI without any significant differences in RFS : this may have been because the RSI questionnaire included the gastroesophageal reflux symptom item.

A Case of Relapsing Polychondritis / 대한피부과학회지

Relapsing polychondritis is a rare inflammatory disorder affecting the cartilaginous structures throughout the body. Although there is no pathognomonic laboratory tests, clinical features, when coupled with histopathologic findings, enable one to make a diagnosis of relapsing polychondritis. A 26-year-old man presented with purplish, swollen, and tender auricles and injected eyes. His symptoms had developed eight months ago and fluctuated irrespective of various medications such as corticosteroids and antibiotics. Histopathologic examination revealed loss of basophilic staining of the cartilage with some dissolution of its structures. Treatment was initiated with dapsone, However, the remission was too immediate to be considered as a effect of dapsone.

Successful Desensitization of A Case of Anaphylaxis to Insulin / 대한피부과학회지

A small proportion of diabetics treated with exogenous insulin, reportedly ranging from 0.1% to 2%, experience systemic allergic reactions to insulin. We saw a 55-year-old male patient with insulin-dependent diabetes mellitus who had developed an anaphylatic reaction immediately after subcutaneous injections of neutral protamine Hagedorn (NPH) human recombinant insulin. Skin tests with insulins and a passive transfer test demonstrated that his reaction was IgE mediated. Since there were no therapeutic alternatives, desensitization with insulin was done following a slow protocol. It was successful and he was able to resume insulin therapy. During two year follow-up period, he did not developed any adverse reaction to insulin.

Multifocal ischemic lesions and focal hematoma formation in a meningitis

Cerebrovascular complications are of the most frequent intracranial complications of bacterial meningitis. Most of the previous reports suggest that the prognosis for the pafients with cerebrovascular complications was unfavorable. We recently experienced a case of meningococcal meningitis with fulminant meningococcemia associated with multifocal non-enhancing lesions on, initial brain MRI. These lesions were hyperintense on T2weighted image and were located in left basal ganglia, both medial thalami, periventricular white matter, left cerebellar hemisphere, and right midbrain and were considered to be resulted from small vessel involvement. Gram negative diplococci were detected by Gram staining of specimens from skin lesion. After antimicrobial therapy and glucocorticoid replacement the patient was recovered without any neurologic sequelae. After one month, follow-up MRI showed resolution of all the ischemic lesions except in midbrain. Additionally there was a small focal hemtoma formation in left basal ganglia. The small hematoma was considered to be resulted from rupture of microaneurysm and disappeared on follow up MRI performed after 3 months. This case suggests that the cerebrovascular complications in meningococcal mningitis might be treated successfully.

Anaerobic Bacterial Isolation in Patients with Chronic Prostatitis Syndrome / 대한비뇨기과학회지

In a general way, anaerobic isolation is troublesome and needs competent personnel and fittings. In addition, bacterial isolation from the prostate is disappointing because of difficulty in interpretation of the results. In this experiment, we tried the best way to isolate anaerobes from the prostate in terms of processing of the specimens such as catching, transportation, etc. We performed this antegrade experiment for 12 months in 1992 and got the results from 43 patients with chronic prostatitis syndrome as follows. l. Age distribution was in broad range between 20 and 54 showing peak incidence in 31-40 years(49% ) and the next in 20-30 years( 30%). 2. Subjective symptoms and signs consisted of perineal discomfort, suprapubic discomfort, frequency, urethral discomfort, dysuria, morning drop, testicular discomfort, and hemospermia 3. Majority of the cases( 36 cases. 83.7% ) were normal in the microscopy of VB1 EPS of 31 cases(72%) showed WBC more than 10/HPF. VB3 of 23 cases(53.5% ) showed than l0/HPF 4. A total of 40 cases showed aerobes in EPS and/or VB3 by culture. However, only 8 cases showed aerobes in EPS and/or VB, exclusive of 32 cases in which aerobes also appeared in VB1. 5. Anaerobic bacteria were cultured only from EPS for a total of 8 cases. There were 2 cases with Bacteroides species, 2 cases with Prevotella bivia, 2 cases with Peptostreptococcus anaerobrus,2 cases with Actinomyces meyeri, 1 case with Eubacterium lentum and 1 case with Eubacterium limosum.

Prognostic Significance of Deoxyribonucleic acid in Stage T1 Bladder Carcinoma / 대한비뇨기과학회지

Flow cytometric nuclear deoxyribonucleic acid (DNA) ploidy analysis was done successfully on 35 specimens of stage T1 bladder carcinoma treated with Bacillus Calmette-Guerin(BCG) between June 1986 and June 1993. The histologic tumor grading was done by the same pathologist. Of the specimens 16(46%) were DNA diploidy, 14(40%) were aneuploidy and 5(14%) were tetraploidy. In the relationship between tumor grade and ploidy, diploidy was seen in all (100%) of 5 patients with grade 1, in 13(48%) of 27 patients with grade 2. On the other hand, non-diploidy in 14(52%) of 27 patients with grade 2, and in all(100%) of 5 patients with grade 3. Mean followup period was 29 months(12-97 months). None of the patients died of tumor during the followup period. None(0%) of 16 patients with diploidy had tumor recurrence, in contrast 8(42%) of 19 patients with non-diploidy tumors recurred(p<0.05). None of the 16 patients with diploidy had progression. but 5(36%) in 14 patients with aneuploidy (5 patients(26%) in 19 patients with non-diploidy) progressed to an advanced stage(p<0.05). The DNA ploidy analysis appears to significantly contribute in the elucidation of possible future recurrence and progression in patients with stage T1 bladder carcinoma.

Factors Affecting the Morbidity after Transurethral Prostatectomy for Benign Prostatic Hyperplasia / 대한비뇨기과학회지

Medical records of 570 patients underwent transurethral prostatectomy from 1985 to 1991 were reviewed to analyze the factors affected morbidity after transurethral prostatectomy retrospectively. Transurethral prostatectomy constituted 16.9% of the urologic operation during above periods and there were benign disease in 513 patients(90%), and carcinoma in 57 patients(10%). Age of the patients ranged from 48 to 93 years old with a mean age of 68.3. Of the patients, 298 patients (52.3%) had significant pre-existing medical problems. The resected net tissue weight ranged from 3 to 73 grams, with a mean of 11.8 grams. The mean operation time was 53 minutes with a range from 25 minutes to 126 minutes. The mortality was 0.36% and the operative morbidity was 20.3%. Increased morbidity was found in patients with a resection time of more than 90 minutes, used glycine of more than 20,000ml and resected weight of more than 31 grams (P<0.05) But age greater than 80 years, azotemia, urinary tract infection, serum Na+ change and pre-existing disease did not increased the operative morbidity. Voiding symptom was improved in 551 patients(96.7%) but in 19 patients(3.3%). persistent voiding difficulty was found. postoperatively at three months The causes of voiding difficulty were as follows ; 10 incontinence patients, 8 detrusor areflexia patients and 1 urethral stricture patient. In conclusion, transurethral prostatectomy is one of the most common operations today with a very low mortality and the overall postoperative results are excellent. Further studies, such as a randomized trial, will be necessary to evaluate the long-term outcome of transurethral prostatectomy further.