OBJECTIVE: Alendronate, a widely used bisphosphonates, acts to inhibit bone resorption by interfering with the activity of osteoclasts. Recently, it has been reported that alendronate also may increase bone proliferation and osteoblastic differentiation. However, little is known about mechanism of the action of alendronate on osteoblast differentiation, especially in transcription level. Inhibitors of DNA binding/ differentiation (Ids) are helix-loop-helix (HLH) transcription factors and play an important role in BMP-induced osteoblast lineage-specific differentiation. Therefore, this study was aimed to investigate the effect of alendronate on osteoblast differentiation and expression of Id-1 and Id-2. METHODS: MC3T3-E1, pre-osteoblast cell line, were treated with alendronate of various concentrations (10(-9) M-10(-4) M) and time periods (24, 48 and 72 hours). And then, the effect of alendronate on osteoblast differentiation was examined by alkaline phosphatase (ALP) activity and RT-PCR for osteoblast differentiation markers such as ALP, type 1 collagen (Col 1), and osteocalcin (OCN). The expressions of Id-1 and Id-2 were measured by RT-PCR. RESULTS: Alendronate treatment increased not only ALP activity, but also expressions of ALP, Col 1, and OCN. Also, alendronate treatment up-regulated the mRNA levels of Id-1 and Id-2 genes. This alendronate-induced osteoblastic differentiation is more effective in lower doses rather than high doses. CONCLUSION: This study shows that the expression of transcription factor Id-1 and Id-2 was increased in a dose-dependent manner during alendronate-induced osteoblast differentiation.
Alendronate ; Alkaline Phosphatase ; Antigens, Differentiation ; Bone Resorption ; Cell Differentiation ; Cell Line ; Collagen Type I ; Delta Sleep-Inducing Peptide ; Diphosphonates ; DNA ; Osteoblasts ; Osteocalcin ; Osteoclasts ; RNA, Messenger ; Transcription Factors

BACKGROUND: Clostridium difficile is one of the most important pathogens responsible for nosocomial diarrhea. Recently, we have frequently experienced culture positive, toxin A enzyme immunoassay negative strains. Therefore, we evaluated the strains with several PCR primer sets to characterize them. METHODS: A total of 351 stool specimens were examined for toxin A using enzyme linked fluorescent immunoassay (ELFA) and also cultured for C. difficile using cycloserine cefoxitine fructose agar incubated under anaerobic conditions. Spore stain and Vitek ANA identification card (BioMerieux, France) were used for identification of C. difficile. We amplified toxin A and toxin B genes in 81 isolates using primers NK1- NK2, NK3-NK2, NK9- NK11, and NK104-NK105. RESULTS: The concordance rate between ELFA and culture was 65.2% (229/351). PCR for the toxin A gene using NK1-NK2, NK3-NK2 and for the toxin B gene using NK104-NK105 showed almost the same results. However, toxin A gene PCR using NK9-NK11 showed that 45.7% (37/81) of the evaluated strains were toxin A (-)/ toxin B(+) variant strains; thus, the corrected sensitivity and specificity of the ELFA based on the PCR results for toxin A and B genes were 65.6% and 100%, respectively. CONCLUSIONS: The low sensitivity of the ELFA results for toxin A was due to the toxin A(-)/toxin B(+) variants of C. difficile, suggesting that the prevalence of the variant strains could be higher in Korea than was expected.
Agar ; Cefoxitin ; Clostridium difficile* ; Clostridium* ; Cycloserine ; Diarrhea ; Fructose ; Genes, vif ; Immunoassay ; Immunoenzyme Techniques ; Korea ; Polymerase Chain Reaction ; Prevalence ; Sensitivity and Specificity ; Spores

BACKGROUND: ChromID Clostridium difficile agar (IDCd; bioMerieux SA, France) is a recently developed chromogenic medium for rapid and specific isolation of C. difficile. We compared the performance of IDCd with that of Clostridium difficile Selective Agar (CDSA). METHODS: A total of 530 fresh stool specimens were collected from patients with clinical signs compatible with C. difficile infection, and cultures for C. difficile were performed on IDCd and CDSA. C. difficile colonies were identified by spore staining, odor, use of an ANI identification test kit (bioMerieux SA), and multiplex PCR for tcdA, tcdB, and tpi. RESULTS: The concordance rate between IDCd and CDSA was 90.6% (480/530). The positivity rates on IDCd on days 1 and 2 (55.6% and 85.0%, respectively) were significantly higher than those on CDSA (19.4% and 75.6%, respectively) (P<0.001 for day 1 and P=0.02 for day 2), but the detection rates on IDCd and CDSA on day 3 were not different (89.4% vs. 82.8%, P=0.0914). On day 3, the recovery rates for non-C. difficile isolates on IDCd and CDSA were 30.2% (160/530) and 22.1% (117/530), respectively (P=0.0075). Clostridium spp. other than C. difficile were the most prevalent non-C. difficile isolates on both media. CONCLUSIONS: The culture positivity rates on IDCd and CDSA were not different on day 3 but IDCd may allow for rapid and sensitive detection of C. difficile within 2 days of cultivation.
Agar/*chemistry ; Bacterial Proteins/genetics ; Bacterial Toxins/genetics ; Clostridium difficile/genetics/*isolation & purification ; DNA, Bacterial/analysis ; Enterocolitis, Pseudomembranous/diagnosis/microbiology ; Enterotoxins/genetics ; Feces/*microbiology ; Humans ; Multiplex Polymerase Chain Reaction ; Reagent Kits, Diagnostic ; Triose-Phosphate Isomerase/genetics

BACKGROUND: Clostidium difficile is one of the most important pathogens responsible for nosocomial diarrhea; therefore, we compared the efficacy of laboratory tests for diagnosing C. difficile diarrhea. METHODS: We evaluated 107 stool specimens using a latex agglutination test (LA) (BD CDT, Culturette CDT, Becton, Dickison and Company, USA) and an enzyme linked fluorescent immunoassay (ELFA) (VIDAS C. difficile Toxin A II, Bio-Merieux sa, Marcy-l'Etoile, France). Stool specimens were cultured using cycloserine cefoxitine fructose agar in anaerobic condition. For identification of C. difficile, spore stain and Vitek ANA identification card (Bio-Merieux sa) were used. Toxin A and toxin B genes were analysed by PCRs using primers NK3-NK2 and NK104N-K105 respectively. RESULTS: The concordance rate between LA and ELFA was 68.2%. Based on the culture results, the sensitivity/specificity of LA and ELFA were 54.8%/100% and 17.8%/100%, respectively. The positive rates of toxin A and B genes were both 90.4%(66/73). Based on the results of PCR assays for toxin A and B genes, the sensitivity/specificity of LA and ELFA were 37.9%/85.7% and 19.7%/100%, respectively. CONCLUSION: Based on C. difficile culture and toxin A and B gene PCR results, the sensitivity of LA was apparently higher than that of ELFA. However, it should not be simply estimated that ELFA has lower capability for detecting toxin A of C. difficile because the possibility of emerging variant strains of C. difficile could not be ruled out. The prevalence of toxigenic strains of C. difficile including variant strains should be studied in Korea.
Agar ; Agglutination* ; Cefoxitin ; Clostridium difficile* ; Clostridium* ; Cycloserine ; Diarrhea ; Fluorescence* ; Fructose ; Immunoassay ; Immunoenzyme Techniques ; Korea ; Latex Fixation Tests ; Latex* ; Polymerase Chain Reaction* ; Prevalence ; Spores

OBJECTIVE: To evaluate the effect of short coasting, by withdrawing both gonadotropins and GnRH agonist (GnRHa), on the prevention in severe ovarian hyperstimulation syndrome (OHSS) without compromising pregnancy outcome. METHOD: Thirty-seven women who had been coasted during COH for IVF were coasted when > or =20 follicles > 15 mm with serum E2 level of 4,000 pg/ml were detected. Coasting was initiated for one or two days depending on the status of follicle on ultrasound and serum E2 level. Both gonadotropin and GnRHa were withheld for coasting. Retrospective study was carried and changes of serum E2 levels, number of oocytes retrieved, fertilization rate, pregnancy rate were compared and analyzed. RESULTS: The mean serum E2 level fell from 6,993 pg/ml on the onset of coasting to 3,396 pg/ml on the day of hCG administration. The mean number of oocytes retrieved and fertilization rate were 15.7 and 70.0%, respectively. Fifteen patients were pregnant (40.6%) and implantation rate was 15.2%. Twenty-six (70.3%) patients were coasted for one day and 11 (29.7%) were coasted for two days. The mean decrease rate of serum E2 level was 43% in one day coasting group and 15% (1st day) and 81% (2nd day) in two day coasting group. The pregnancy outcome was similar between the two groups. After coasting, no severe or moderate OHSS occurred in any patients and mild OHSS occurred in 3 (8.1%) patients. CONCLUSIONS: Coasting for one or two days can be used successfully in the prevention of OHSS without compromising IVF cycle outcome.
Female ; Fertilization ; Gonadotropin-Releasing Hormone* ; Gonadotropins* ; Humans ; Oocytes ; Ovarian Hyperstimulation Syndrome* ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies ; Ultrasonography

BACKGROUND: Korean national cancer screening program selected fecal occult blood test (FOBT) as a primary screening method of colorectal carcinoma in adult > or =50 yr old irrespective of symptom. Notice to pre-analytical errors is especially important for the FOBT because examinees collect and submit their specimens to laboratories by themselves. We examined the influences of the fecal storage temperatures, durations and with or without buffer on the FOBT results. METHODS: Thirty FOBT-positive specimens above 100 ng/mL were used for the study from July to August 2008. Quantitative FOBT was performed with OC-sensors II (Eiken Chemical Co., Japan). Each specimen was divided into 4 groups. Two groups in plastic buffer-free containers were kept either at 4degrees C or room temperature (25-28degrees C), respectively. Another two groups in buffer-tubes were also kept either at 4degrees C or room temperature. Each group was repeatedly examined with same method every 24 hr up to 120 hr. RESULTS: Eleven specimens (36.7%) in buffer-free containers converted to negative results (below the 100 ng/mL) after 24 hr and 17 specimens (56.7%) did after 48 hr at room temperature. Ten specimens (33.3%) in buffer-free containers converted to negative after 48 hr at 4degrees C. Specimens contained in buffer-tubes showed little change; 3 specimens (10.0%) at room temperature and no specimen at 4degrees C showed negative conversions after 48 hr. CONCLUSIONS: Buffer-tube minimizes false negative FOBT results during pre-analytical delay of specimen. The examinees using buffer-free containers need to be educated to hand in their specimens to laboratories as soon as possible.
Buffers ; Colorectal Neoplasms/*diagnosis ; Humans ; Middle Aged ; *Occult Blood ; *Specimen Handling ; Temperature ; Time Factors

OBJECTIVE: To evaluate the effects of serum E2 levels on the day of hCG administration on the pregnancy outcome of IVF-ET after controlled ovarian hyperstimulation (COH). METHODS: Data of 455 cycles of fresh IVF-ET with COH were retrospectively investigated. Serum E2 levels on the day of hCG administration were categorized into 5 groups; A (<1,000 pg/mL), group B (1,000~2,000 pg/mL), group C (2,000~3,000 pg/mL), group D (3,000~4,000 pg/mL), and group E (> or =4,000 pg/mL). RESULTS: Mean E2 levels on the day of hCG administration were 3,745.3 pg/mL and mean number of retrieved oocytes were 10.1. Of 455 cycles, 148 (32.5%) cycles were clinically pregnant. Implantation rate was 12.2% and delivery rate was 18.7%. The number of obtained oocytes increased with increasing levels of serum E2. Pregnancy rate gradually increased as E2 levels increased up to the group D, but began to fall in the group E. In younger women (<38 yrs), the IVF-ET outcomes were similar to those of total patients but in older women (> or =38 yrs), pregnancy rate and delivery rate were significantly higher in the group C than other groups. CONCLUSIONS: This result shows that serum E2 levels have a concentration-dependent effect on the pregnancy outcome and there is an optimal range of E2 levels to achieve for a successful pregnancy. Excessive E2 levels seem more deleterious to the pregnancy outcome in older women aged > or =38 years.
Aged ; Female ; Humans ; Oocytes ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies

OBJECTIVE: Ovarian angiogenesis plays an important role in folliculogenesis. However, little is known about the expression of angiogenic factors during follicular development according to female age. Stromal cell derived factor-1alpha (SDF-1alpha) plays a role in granulosa cell survival and embryo quality as an angiogenic chemokine. Leptin is also involved in folliculogenesis and angiogenesis. This study examined expression of SDF-1alpha and leptin, and their effects on the expression of angiogenic factors in the ovary during follicular development according to female age. METHODS: Ovaries were collected from C57BL mice of two age groups (6-9 weeks and 24-26 weeks) at 6, 12, 24, and 48 hours after 5 IU pregnant mare's serum gonadotropin (PMSG) injection. The expression of ovarian SDF-1alpha and leptin mRNA was evaluated by RT-PCR. In the organ culture experiment, the ovaries were cultured in transwell permeable supports with Waymouth's medium treated with various doses of SDF-1alpha (50-200 ng/mL) or leptin (0.01-1 microg/mL) for 7 days. Then, mRNA expression of vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), and visfatin were examined in the cultured ovaries. RESULTS: Expression of SDF-1alpha and leptin in the ovary was significantly lower in the aged mouse group compared to the young mouse group (p<0.05). Expression of these two factors increased with follicular development after PMSG administration. SDF-1alpha treatment stimulated visfatin expression in a dose-dependent manner, while leptin treatment significantly increased eNOS expression. CONCLUSION: These results suggest that decrease of ovarian SDF-1alpha and leptin expression may be associated with aging-related reduction of ovarian function. SDF-1alpha and leptin may play a role in follicular development by regulating the expression of angiogenic factors in mouse ovaries.
Aged ; Aging ; Angiogenesis Inducing Agents ; Animals ; Chemokine CXCL12 ; Embryonic Structures ; Female ; Gonadotropins ; Granulosa Cells ; Humans ; Leptin ; Mice ; Mice, Inbred C57BL ; Nicotinamide Phosphoribosyltransferase ; Nitric Oxide Synthase Type III ; Organ Culture Techniques ; Ovary ; RNA, Messenger ; Stromal Cells ; Vascular Endothelial Growth Factor A

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Databases, Chemical ; Subject Headings

Premature ovarian failure (POF) is a syndrome defined as hypergonadotropic hypogonadism associated with amenorrhea, oligomenorrhea or other forms of menstrual irregularity for at least 3 consecutive months before the age of 40. The management of POF is approached by HRT, emotional support and infertility treatment. Women with premature ovarian failure who desire to become pregnant are best treated by assisted reproductive technology with donor oocyte. However, POF has the possibility of a 5-10% spontaneous pregnancy. The physician should recommend the patient to consult with their physician if they have any symptoms of pregnancy or no withdrawal bleeding after HRT. Therefore we report two cases of spontaneous pregnancies in women with premature ovarian failure.
Amenorrhea ; Female ; Hemorrhage ; Humans ; Hypogonadism ; Infertility ; Oligomenorrhea ; Oocytes ; Pregnancy* ; Primary Ovarian Insufficiency* ; Reproductive Techniques, Assisted ; Tissue Donors