Tinea incognito (TI) is a dermatophytic infection which has lost its typical clinical appearance because of improper use of steroids or calcineurin inhibitors. The incidence of TI is increasing nowadays. We conducted retrospective review on 283 patients with TI from 25 dermatology training hospitals in Korea from 2002-2010 to investigate the demographical, clinical, and mycological characteristics of TI, and to determine the associated risk factors. More than half (59.3%) patients were previously treated by non-dermatologists or self-treated. The mean duration of TI was 15.0 +/- 25.3 months. The most common clinical manifestations were eczema-like lesion, psoriasis-like, and lupus erythematosus-like lesion. The trunk and face were frequently involved, and 91 patients (32.2%) also had coexisting fungal infections. Among 67 isolated strains, Trichophyton rubrum was the most frequently detected (73.1%). This is the largest study of TI reported to date and the first investigational report concerning TI in Korea. We suggest that doctors should consider TI when a patient has intractable eczema-like lesions accompanied by tinea pedis/unguium. Furthermore, there should be a policy change, which would make over-the-counter high-potency topical steroids less accessible in some countries, including Korea.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Demography ; Eczema/pathology ; Face/pathology ; Female ; Humans ; Lupus Erythematosus, Cutaneous/pathology ; Male ; Middle Aged ; Psoriasis/pathology ; Republic of Korea ; Retrospective Studies ; Risk Factors ; Tinea/*diagnosis/microbiology ; Trichophyton/isolation & purification ; Young Adult

OBJECTIVE: The purpose of this study was to examine in vitro development of early preimplantation mouse embryos in various kind of serum-free conditioned media (SF-VCM) manufactured from DMEM cultured with Vero Cells. METHODS: A total of 846 two-cell mouse embryos were cultured in different kind of SF-VCM. SF-VCMs were divided into SF-VCM-10, -30 and -50 by media volume using DMEM #1 media, and divided into SF-VCM #1, #2 and #3 by controlled concentration of glucose and pyruvate (manufactured by DMEM #1: mixed three volume of DMEM-G (DMEM with glutamine without glucose and pyruvate) and one volume of DMEM-GGP (DMEM with glutamine, glucose, pyruvate), #2: mixed same volume of DMEM-G and DMEM-GGP and #3: mixed one volume of DMEM-G and three volume of DMEM-GGP, respectively). Experimental groups were mainly added 10% SSS, and 20% hFF was added to only Control group co-cultured with Vero cells. Development of embryos was observed every 24 hours. Results between different groups were analyzed using Chi-square test, and considered statistically significant when P-value was less than 0.05. RESULTS: In vitro developmental rate by each cleavage stages of mouse embryos cultured in SF-VCMs with a various volumes were significantly (P<0.05) higher in SF-VCM-30 (morula< or =: 97.2%, Blastocyst (BL)< or =: 97.2%, Hatching BL< or =: 82.2%) than other groups. In the rate of development on in vitro co-culture vs. a various SF-VCMs manufactured by DMEM controlled concentration of glucose and pyruvate, Group I (SF-VCM #1) was higher than other groups in each cleavage stages (morula< or =: 98.1%, Blastocyst (BL)< or =: 97.1%, hatching BL< or =: 81.7%, respectively). Moreover, specially, in the developmental rate into the hatching blastocyst < or = after 96 hours in vitro culture, Group I (81.7%) was significantly higher than control group (67.6%, P<0.05). CONCLUSION: SF-VCM #1 manufactured by volume of 30 mL DMEM #1 media cultured in vitro for 48 hours in 250 mL flask was the most effective on in vitro developmental rate of mouse preimplantation embryos. Therefore, it is expected that SF-VCM #1 has application to human IVF-ET.
Animals ; Blastocyst ; Coculture Techniques ; Culture Media, Conditioned ; Embryonic Structures ; Fertilization in Vitro ; Glucose ; Glutamine ; Humans ; Mice ; Pyruvic Acid ; Vero Cells

OBJECTIVE: The purpose of this study was to examine the effects of vero cells co-culture system on the in vitro development of mouse preimplantation embryos in culture media with different composition of glucose and pyruvate. METHODS: Two-cell embryos were collected from 4-5 weeks old ICR mice. Seven hundreds twenty two embryos were cultured with or without vero cells monolayer in four media with different compositions that was manufactured by two DMEM media with (DMEM-GGP) or without (DMEM-G) glucose and pyruvate. In control, DMEM-G medium which is currently using for human embryo culture in our infertility clinic was used. Group I (DMEM-G(1/4)GP) was cultured in medium which was mixed three volume of DMEM-G and one volume of DMEM-GGP, and group II (DMEM-G(1/2)GP) was cultured in medium which was mixed same volume of DMEM-G and DMEM-GGP, and group III was cultured in DMEM-GGP. All media were added to 20% hFF. Results between different groups were analyzed using a Chi-square test, and considered statistically significant when p value was less than 0.05. RESULTS: The developmental rate into 3-cell Animals ; Blastocyst* ; Coculture Techniques* ; Culture Media ; Embryonic Development ; Embryonic Structures ; Female ; Glucose* ; Humans ; Infertility ; Mice* ; Mice, Inbred ICR ; Pregnancy ; Pyruvic Acid* ; Vero Cells*

OBJECTIVE: Mammalian embryos undergo changes of energy environment for transfer from oviduct to uterus. An experimental design was used to examine the effects of glucose on the development in vitro of mouse embryos. METHODS: Two cell embryos were recovered from ICR female mice (3-4 weeks) at 46-50 hrs after 5 IU hCG injection (mated just after hCG injection) and cultured in 50 micro gram MEM droplets supplemented with nothing (control; n=46), 0.5 mM glucose (Group A; n=46) or 3.15 mM glucose (Group B; n=46) under mineral oil. All experimental media were supplemented with 20% human follicular fluid. Results were observed: (i) the number of zona-intact blastocysts (ZiB); (ii) the number of zona-escaped blastocysts (ZeB; hatching~hatched); (iii) the mean cell numbers; and (iv) the proportion of inncer cell mass (% ICM) in the blastocysts. RESULTS: Total blastocyst formation rates were (NS) in glucose groups (group A: 52.2; B: 47.8%) than control group (60.9%). ZiB rates the highest (p<0.05) in control (47.8%) than those in group A (21.7%) and B (28.3%). ZeB rates the highest (NS) in group A (30.4%) than those in control (13.0%) and group B (19.6%). Blastocysts, cultured in group B (50.5), had the highest (NS) mean cell numbers compared with the others (control: 39.2; group A: 45.6). The % ICM in blastocysts cultured in group A (20.6%) was the highest (NS) than those of other tested groups (control: 15.2%; group B: 13.9%). CONCLUSION: This study shows that a low dose of glucose added to culture medium increases the developmental capacity of 2 cell embryos in mice.
Animals ; Blastocyst* ; Cell Count* ; Embryo, Mammalian ; Embryonic Structures* ; Female ; Follicular Fluid ; Glucose* ; Humans ; Mice* ; Mineral Oil ; Oviducts ; Research Design ; Uterus

OBJECTIVE: This present study was conducted to examine the effects on in vitro growth of pre-antral mouse follicles by thecal-stromal cells attached to around the follicular membrane in culture medium without hormones. METHODS: Pre-antral follicles (100-130 micro meter) used in our studies were isolated mechanically by fine 30 G needles attached to 1 ml insulin syringe from mice ovaries of 20-25 days old female ICR strain. Isolated pre-antral follicles were divided into three groups by attached status of thecal-stromal cell layers to follicular membrane. Follicular Initial diameters of group I was 112.5 (mean) +/- 6.2 (SD) micro meter (n=31), group II was 112.8 +/- 7.8 micro meter (n=23), and group III was 115.1 +/- 6.5 micro meter (n=27). Divided pre-antral follicles were washed in fresh Ham's F-10 medium and cultured in 20 micro liter droplets of DMEM with glutamine, glucose and pyruvate under mineral oil on the 60 mm culture dish. All experimental media were supplemented with 10% FBS without hormones. Media were exchanged wholly by fresh media every 2 days in culture. Diameters of intrafollicular oocytes and cultured pre-antral follicles were measured using an precalibrated cross ocular micrometer at X200 every day during 6 days in vitro culture. Results were considered statistically significant when p value was less than 0.05 using Student's t-test. RESULTS: The mean and standard deviation (SD) of initial diameters of pre-antral follicles was 113.5 +/- 2.2 micro meter in three groups. Follicular growth rate of group III was significantly (p<0.05) higher in whole culture periods than group I and group II. Rate of follicular growth between group I and group II were not significant difference in culture for 1 to 3 days. However, group II was significantly higher than group I in culture for 4 days. Diameters of grown follicles for 6 days was 155.2 +/- 18.7 micro meter, 196.9 +/- 24.1 micro meter and 284.2 +/- 47.6 micro meter in group I, group II and group III, respectively. Growth rate of intrafollicular oocytes between three groups were not difference and revealed to continuous growth patterns in whole culture periods. Follicular diameter after culture for 7 days were not measured because of disruption of follicular structure or outgrowth of granulosa cells. CONCLUSION: Pre-antral follicles were grown very well in DMEM medium without hormones. Thecal- stromal cells attached to the surface of follicular membrane has acted effectively in vitro growth of follicles.
Animals ; Female ; Glucose ; Glutamine ; Granulosa Cells ; Humans ; Insulin ; Membranes ; Mice* ; Mineral Oil ; Needles ; Oocytes ; Ovary ; Pyruvic Acid ; Stromal Cells ; Syringes

OBJECTIVE: This study was performed to evaluate and compare the embryonic developmental capacity and pregnancy rates in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with ejaculated sperm or testicular sperm cycles. MATERIALS AND METHODS: Fertilization was examined in the following morning after IVF (group I), ICSI (group II) or TESE-ICSI cycles (group III). Fertilized oocytes were co-cultured with Vero cells until embryo transfer (ET). On day 2 and 5~7, grades of embryos (<4- or > or =4-cell) and blastocysts (BG1, 2, 3 or early) were evaluated. Clinical pregnancy rate was determined by detecting G-sac with transvaginal ultrasonogram. We analyzed the results bychi2 and Student's t-test and considered statistically significant when P value was less than 0.05. RESULTS: Fertilization rate was significantly higher (p<0.05) in group I (79.0+/-21.2%) than in group II and III (56.8+/-21.6% and 36.7+/-25.3%). Cleavage and blastulation rate of group I (95.8+/-13.8% and 59.5+/-25.3%) were significantly higher (p<0.05) than those of group III (83.4+/-18.6% and 40.4+/- 36.5%). Clinical pregnancy rate was significantly higher (p<0.05) in group I and II (40.7% and 41.7%) than that in group III (12.5%). No differences were found in the rates of multiple pregnancy and abortion among three groups. Embryonic implantation rate was higher in group I (15.1+/-20.2%, p<0.05) and II (14.7+/-20.6%, NS) than that in group III (5.1+/-15.6%). However, embryonic implantation rate was increased in ET with blastocyst(s) among three groups. CONCLUSIONS: Fertilized oocytes obtained from TESE-ICSI were harder to be successfully cultured to blastocyst stage for 5~7 days than that from IVF cycles. However, all blastocyst(s) ET increased the embryonic implantation rate equally in IVF, ICSI and TESE-ICSI cycles.
Blastocyst ; Embryo Transfer ; Embryonic Development* ; Embryonic Structures ; Female ; Fertilization ; Fertilization in Vitro ; Oocytes* ; Pregnancy ; Pregnancy Rate* ; Pregnancy* ; Pregnancy, Multiple ; Sperm Injections, Intracytoplasmic* ; Spermatozoa ; Ultrasonography ; Vero Cells

OBJECTIVE: The purpose of this study was to examine the effects on in vitro development of early preimplantation mouse embryos in DMEM medium with glutamine which was controlled by different composition of glucose and pyruvate. METHODS: Four hundred and nineteen mouse 2-cell embryos were cultured in four different media with different composition of glucose and pyruvate for 96 hours. The DMEM-G contained L-glutamine for energy sources was used for control group. Group I embryos were cultured in the medium that mixed one volume of DMEM-GGP contained L-glutamine, D-glucose and sodium pyruvate for energy sources with three volume of DMEM-G, and group II embryos were cultured in the medium that mixed with same volume of DMEM-G and DMEM-GGP, and group III embryos were cultured in DMEM-GGP. RESULTS: At 24 hours, the development into >or=3-cell was significantly higher (p<0.05) in group I (93.3%) than control (84.6%). The development into >or=8-cell was significantly higher in group I (73.1%) than control (44.2%), group II (59.6%) and III (45.8%), and also group II was significantly higher than control and group III. At 48 hours, the development into >or=morula was significantly higher in group I (90.4%) and II (86.5%) than control (73.0%). However, the development into blastocyst, in group III (15.0%) was significantly lower than control, group I and II. At 72 hours, the development into >or=expanded blastocyst was significantly higher in group I (69.2%) than group III (47.7%), and total blastocyst was significantly higher in group I (80.8%) than control (66.3%) and group III (67.3%). At 96 hours, the development into >or=hatching blastocyst was significantly higher in group I (78.8%) than control (61.5%) and group III (57.9%), and also, total blastocyst was significantly higher in group I (85.6%) than control (69.2%) and group III (72.0%). CONCLUSION: The development of early preimplantation mouse embryos cultured in group I medium that mixed one volume of DMEM-GGP with three volume of DMEM-G was better than other groups during the culture period.
Animals ; Blastocyst* ; Embryonic Structures ; Glucose* ; Glutamine* ; Mice* ; Pyruvic Acid* ; Sodium

Transfer of human embryos to the uterus at the blastocyst stage has several advantages. There may be an improvement in success rates of pregnancy due to better synchronization of the uterine and embryonic development, self-selection of embryos for transfer along with the possibility of reducing the number of embryos transferred and the risk of multiple pregnancies without altering the overall pregnancy rate would be decreased. However, embryos are transferred to the uterus on day 2 or 3 after insemination before the blastocyst stage is reached in many cases. This may be a reflection of sub-optimal culture conditions although inherent abnormalities like chromosomal anomalies will also contribute to the loss.1~8 Physiologically, the human endometrium prepares itself to its optimum approximately on days 5~7 after ovulation so as to receive a cavitated blastocyst from the fallopian tube for successful implantation.1,9,10 However, conventionally, in most programs offering in vitro fertilization (iVF), 4~8 cell stage embryos are replaced to the uterus.1,9,11 This asynchrony of embryonic stage and preparation of endometrium may be one major contributory cause of increased abortion and low take-home baby rates in infertility patients.1,10~12 Leaving all embryos in extended culture until they develop to the blastocyst stage might result in cancellation of the embryo transfer (ET) procedure if none of the embryos reach that stage. This could have a major adverse psychological impact on the patient, and also denies her the possibility of implantation of early embryos that did not develop to the blastocyst stage in vitro but might have done so in vivo.2 To avoid such events while explore the feasibility of blastocyst transfer, subsequent ET (SET) of early embryos and blastocysts was applied on day 2~3 and 5.2,6,7,13,14 The current study was conducted to investigate the effectiveness of attempted SET of blastocysts on day 5~7 following initial multi-cell embryos transfer on day 2, 3 or 4 in iVF cycles.
Blastocyst ; Embryo Transfer* ; Embryonic Development ; Embryonic Structures* ; Endometrium ; Fallopian Tubes ; Female ; Fertilization in Vitro ; Humans ; Infertility ; Insemination ; Ovulation ; Pregnancy ; Pregnancy Rate ; Pregnancy, Multiple ; Uterus

OBJECTIVE: To evaluate whether co-culture of oocytes on vero cell monolayers from Day 0 (Day 0 group) after egg retrieval results in an increase in developmental capacity such as fertilization rate, embryo quality, blastulation and clinical pregnancy rate compared with co-culture of oocytes from Day 1 (Day 1 group). METHODS: Sperms were treated with Hams F-10 supplemented with 10% human follicular fluid (hFF). Vero cells for co-culture were prepared in TCM-199 with 10% FBS. Oocytes were co-cultured from Day 0 and fertilized oocytes were co-cultured from Day 1 on vero cell monolayers in DMEM with 10% and 20% hFF, respectively after egg retrieval. On day 1, 2 and 5, fertilization rate and grade of embryos and blastocysts were evaluated. Results (fertilization rate, cleavage rate, grade of embryos and blastocysts and pregnancy rate) were considered statistically significant when p value was less than 0.05 using t-test and ci2. RESULTS: In sibling oocytes of same cycles, no differences were found in fertilization rate (94.6 vs. 91.4%), cleavage rates (94.6 vs. 91.4%), embryo grade (on day 2 and 3) and blastulation (65.6 vs. 57.0%) and their grade. In different oocytes of different cycles (patients), no differences were found in fertilization (79.8 vs. 78.3%), cleavage rates (77.7 vs. 76.4%) and blastulation (56.0 vs. 45.3%), but pregnancy rate was higher in the Day 0 group than in the Day 1 group (60.0 vs. 42.9%). CONCLUSIONS: This study revealed that the embryonic development capacities were not affected by the different co-culture time in the sibling oocytes of same cycles. Although no statistical significance, because of small size of study, there was a trend for higher pregnancy rates in Day 0 group compared to Day 1 group in different oocytes of different cycles.
Blastocyst ; Coculture Techniques* ; Embryonic Development* ; Embryonic Structures ; Female ; Fertilization ; Follicular Fluid ; Humans ; Oocytes* ; Ovum ; Pregnancy ; Pregnancy Rate ; Siblings ; Spermatozoa ; Vero Cells

OBJECTIVE: The objective of this study was to examine the effect on development of mouse preimplantation embryos in culture media with different composition of energy sources in vitro culture. METHODS: Two hundred and seventy one two-cell embryos were cultured in four different culture system for 96 hours. Group I (n=61) was cultured in DMEM-G (DMEM with glutamine) only, groupII (n=64) was cultured in DMEM-GGP (DMEM with glutamine, glucose and pyruvate) only, group III (n=72) was cultured for 48 hours in DMEM-G and then transferred to DMEM-GGP and group IV (n=74) was cultured for 48 hours in DMEM-GGP and then transferred to DMEM-G. Development of embryos in each group was observed every 24 hours. RESULTS: After 24 hours, the rate of development > or = 3-cell was significantly higher in groupII (87.5%) and IV (86.5%) compared with group I (59.0%) and III (62.5%). After 48 hours, the rate of development into > or = morula stage was significantly higher in GroupII (79.7%) and IV (86.5%) compared with group I (34.4%) and III (37.5%). After 72 hours, the rate of development into blastocyst was significantly higher in group IV (74.3%) compared with group I (49.2%) and III (45.8%). After 96 hours, the rate of development into > or = expanded blastocyst was significantly higher in group IV (70.3%) compared with group I (32.8%),II (53.1%), and group III (40.3%). CONCLUSION: Mouse preimplantation embryos development was the most effective in culture system with DMEM-GGP for 48 hours and then transferred to DMEM-G.
Animals ; Blastocyst* ; Culture Media* ; Embryonic Structures ; Glucose ; Glutamine ; Mice* ; Morula ; Pyruvic Acid