Clinical application of genome-wide single nucleotide polymorphism genotyping and karyomapping for preimplantation genetic testing of Charcot–Marie–Tooth disease

Purpose@#Preimplantation genetic testing for monogenic disorders (PGT-M) has been successfully used to prevent couples with monogenic disorders from passing them on to their child. Charcot–Marie–Tooth Disease (CMT) is a genetic disorder characterized by progressive extremity muscle degeneration and loss of sensory function. For the first time in Korea, we report our experience of applying single nucleotide polymorphism genotyping and karyomapping for PGT-M of CMT disease. @*Materials and Methods@#Prior to clinical PGT-M, preclinical tests were performed using genotypes of affected families to identify informative single-nucleotide polymorphisms associated with mutant alleles. We performed five cycles of in vitro fertilization PGT-M in four couples with CMT1A, CMT2A, and CMT2S in CHA Fertility Center, Seoul Station. @*Results@#From July 2020 through August 2021, five cycles of PGT-M with karyomapping in four cases with CMT1 and CMT2 were analyzed retrospectively. A total of 17 blastocysts were biopsied and 15 embryos were successfully diagnosed (88.2%).Ten out of 15 embryos were diagnosed as unaffected (66.7%). Five cycles of PGT-M resulted in four transfer cycles, in which four embryos were transferred. Three clinical pregnancies were achieved (75%) and the prenatal diagnosis by amniocentesis for all three women confirmed PGT-M of karyomapping. One woman delivered a healthy baby uneventfully and two pregnancies are currently ongoing. @*Conclusion@#This is the first report in Korea on the application of karyomapping in PGT-M for CMT patients. This study shows that karyomapping is an efficient, reliable and accurate diagnostic method for PGT-M in various types of CMT diseases.

Identification of unbalanced complex chromosomal rearrangements in IVF-derived embryos during NGS analysis of preimplantation genetic testing: A case report

Complex chromosome rearrangements (CCRs) are structural chromosomal rearrangements involving at least three chromosomes and more than two breakpoints. CCR carriers are generally phenotypically normal but related to higher risk of recurrent miscarriage and having abnormal offspring with congenital anomalies. However, most of CCR carriers are not aware of their condition until genetic analysis of either abortus or affected baby or parental karyotyping is performed. Herein, we present the case that CCR carrier patients can be identified by preimplantation genetic testing of preimplantation embryos. An infertile male patient with severe oligoasthenoteratozoospermia was diagnosed balanced reciprocal translocation, 46,XY,t(3;11) (p26;p14) at first. After attempting the first preimplantation genetic testing for structural rearrangement (PGT-SR) cycle, we found the recurrent segmental gain or loss on 21q21.3-q22.3 of five out of nine embryos. As a result of karyotype re-analysis, the patient’s karyotype showed a balanced CCR involving chromosomes 3, 11, and 21 with three breakpoints 3p26, 11p14, and 21q21. The patient underwent two PGT-SR cycles, and a pregnancy was established after the transfer of an euploid embryo in the second cycle. Amniocentesis confirmed that the baby carried normal karyotype without mosaicism. At 37 weeks gestation, a healthy girl weighting 3,050 g was born.

Characterization of a prenatally diagnosed de novo der(X)t(X;Y)(q27;q11.23) of fetus

A 31-year-old woman, who was pregnant with twins, underwent chorionic villus sampling because of increased nuchal translucency in one of the fetuses. Cytogenetic analysis showed a normal karyotype in the fetus with increased nuchal translucency. However, the other fetus, with normal nuchal translucency, had a derivative X chromosome (der(X)). For further analysis, fluorescence in situ hybridization (FISH) and additional molecular studies including fragile X analysis were performed. FISH analysis confirmed that the Y chromosome was the origin of extra segment of the der(X). The X-chromosome breakpoint was determined to be at Xq27 by FMR1 CGG repeat analysis, and the Y-chromosome breakpoint was determined to be at Yq11.23 by the Y chromosome microdeletion study. To predict the fetal outcome, the X-inactivation pattern was examined, and it revealed non-random X inactivation of the der(X). To the best of our knowledge, the identification of an unbalanced Xq;Yq translocation at prenatal diagnosis has never been reported. This study was performed to identify precise breakpoints and the X-inactivation pattern as well as to provide the parents with appropriate genetic counseling.

Antitumor immunity induced by tumor cells engineered to express a membrane-bound form of IL-2

Transduction of cytokine gene into tumor cells is a promising method of tumor therapy, but the value is limited by accompanying side effects. To focus antitumor immune response to tumor antigen-specific CTL, we developed an antitumor vaccine by transfecting modified IL-2 gene in a membrane-bound form (mbIL-2) into B16F10 melanoma cells. The mbIL-2 clone showed reduced tumorigenicity and metastatic ability, and inhibited metastasis and prolonged the survival of mice against B16F10 cells. The inhibition of B16F10 metastasis by mbIL-2 was accompanied by the increment of CD8+ T cells. The metastasis of mbIL-2 clone was significantly increased in the CD8+ T cell-depleted mice, but not in CD4+ T cell depleted mice. Spleen cells immunized with the mbIL-2 clone showed higher CTL activity towards B16F10 cells than those immunized with control cells. The size of CD8+ T cell population in the lung of mice injected with the mbIL-2 clone was markedly greater than that of mice injected with B16F10 cells, but there was no detectible change in CD4+ and CD8+ T cell populations of lymph nodes and spleen. These results suggest that when the mbIL-2 clone is introduced into the blood stream, it migrates mainly to lung and activates CD8+ T cells in situ, possibly by direct priming. Such a tumor vaccine may ameliorate the toxic side effects encountered with conventional cytokine gene therapy.

Comparison of estrogen receptor beta expression between breast cancer and normal mammary tissue and relationship with clinicopathological factors / 한국유방암학회지

PURPOSE: To verify the difference of estrogen receptor beta (ER beta) expression between breast cancer and normal mammary tissue and the roles of ER beta in prognosis of breast cancer, its expression was investigated in normal mammary and breast cancer tissues METHODS: The ER beta expression of 89 normal mammary and 100 breast cancer tissues was examined using immunohistochemistry. The staining signal was scored by estimating the proportion (range, 0-5) and intensity scores (range, 0-3) of positive cells. The ER beta expression was considered as positive if the total score (IHC score; range, 0-8) was 3 or more. The ER beta expressions were compared between normal mammary and breast cancer tissues. The association of ER beta expression with other clinicopathological factors was also investigated. The distant relapse free survival (DRFS) and overall survival (OS) rates were compared according to the ER beta expression. RESULTS: Eighty-eight of the 89 (98.9%) cases of normal mammary tissues and 74 of the 89 (83.1%) counterpart breast cancer tissues showed positive staining with decreased ER beta expression in the breast cancer compared to the normal mammary tissue with statistical significance (p=0.026). In breast cancer, the ER beta expression was found to have a positive correlation with the ER expression but with only marginal significance (p=0.063). There was no correlation between the ER beta expression and other clinicopathological factors (age, tumor size, nodal status, histological grade, progesterone receptor status, and HER-2 expression). The 5 year DRFS and OS rates were found to be independent of ER beta expression. CONCLUSION: The ER beta expression was significantly decreased in cancer tissues. Further study with a sufficient number of patients is needed to verify the roles of ER beta during breast cancer carcinogenesis and clinical value.

The effect of growth factor receptor HER2 on the estrogen receptor transcriptional activities and its implications / 한국유방암학회지

PURPOSE: Until recently, breast cancer carcinogenesis has not been fully understood, but the roles of estrogen receptors(ERs) and growth factor receptors(like HER2) were known to be important. Growth factors have been shown to synergize in the E2 signaling pathway, although the actual molecular mechanism remains largely unknown. To investigate the effect of HER2 overexpression on the ERE(estrogen responsive element)-mediated transcriptional activity of the ERs, this study was designed. METHODS: NIH3T3 cells, T6-17 cells (NIH3T3 cells with stably transfected with HER2), and MCF-7 cells were maintained in dextran-coated charcoal stripped 10% Dulbecco's Modified Eagle Medium (DMEM). Transient transfection of constructs (pcDNA3-ER alpha, pcDNA3-ER beta, pERE-luc, pAP-1-luciferase, and pcDNA-HER2) into each cells was performed using the Lipofectamine PLUS(TM) system. Reporter gene assays using ERE-luciferase or AP-1-luciferase were used to measure the ER transcriptional activities after treatment with estradiol (E2) and tamoxifen. RESULTS: Reporter gene assay using ERE-luciferase in both ER alpha and ER beta, showed much less responsiveness to estrogen in HER2 overexpressing T6-17 cells than in NIH3T3 cells, but there was no remarkable difference after treatment with tamoxifen. The AP-1-mediated transcriptional activity was increased in ER beta after tamoxifen treatment, but it disappeared in HER2-expressing T6-17 cells. The responsiveness to estrogen in HER2-transfected MCF-7 cells was also slightly less than in the control MCF-7 cells, and the ERE-mediated transcriptional activity of estrogen in MCF-7 cells was decreased, in a dose-dependent manner, after HER2 transfection. CONCLUSION: Coexpression of HER2 and ER seems to make cells less responsive to estrogen stimulation, and decrease the ERE-mediated transcriptional activity in both ER alpha and ERbeta. These results suggest that the expression of HER2 reduces the estrogen dependency in cell growth and eventually induces estrogen independent-growth.

Clinical Features of Spontaneous Neonatal Gastric Perforation

PURPOSE: Spontaneous neonatal gastric perforation is a rare neonatal event which is associated with high mortality. The aim of this retrospective study is to investigate the diagnosis and the treatment of neonatal spontaneous gastric perforation. METHODS: From January 1991 to December 2000, eight cases of neonatal gastric perforation were presented at Keimyung University Dongsan Medical Center. RESULTS: Among 8 neonates (7 males and 1 females), 2 were premature infants. 7 cases (87.5%) had symptoms onset in first 4 days of life. The most presenting signs were abdominal distension, vomiting, and respiratory difficulty. The most common X-ray finding was pneumoperitoneum (87.5%) and the most common site of perforation was greater curvature (75%). The combined congenital anomalies were malrotation of small intestine, teratoma, and partial duodenal obstruction. The predisposing causes may be associated with congenital intestinal anomaly, necrotizing enterocolitis, and too fast increased formula. All the cases underwent surgical repairs. The mortality rate was 37.5%, and the surgical procedure performed 24 hours after presumed disease onset represented poor outcome (mortality rate 67%). CONCLUSION: This report suggests that early diagnosis and early management before clinical deterioration of metabolic status may improve the prognosis for neonatal spontaneous gastric perforation patient.

Characteristics of the regimens for patients with pulmonary tuberculosis registered at public health centers in Seoul

BACKGROUND: Through the control of tuberculosis at 22 public health centers under the National Tuberculosis Control Program, this study is purposed to examine the situation of the tuberculous patients and the characteristics of the therapeutic regimens. METHODS: The data was obtained from 8091 medical records of pulmonary tuberculous patients who were registered for treatment at public health center in Seoul during the year of 1993. It was comparatively analysed by the general characteristics(gender, age, chest X-ray findings, sputum results, treatment results, side effects, combined diseases and accompanied extra-pulmonary tuberculosis) according to various regimens of the tuberculosis. RESULTS: The male patients were 5144, the female were 2947. 34.1% of patients were between 21 and 30years of age. Short course regimen was 97.1% and long course regimen was 2.9%. According to chest X-ray findings minimal 53.5%, moderately advanced 41.2%, far advanced 5.3%. Sputum AFB negative was 52.2% and positive was 47.8%. Therapeutic efficiency was high in short course regimen. Among the side effects, dermatologic problems was high and at the regimen of EHRZ side effects were developed highly. Combined diseases were liver diseases(5.2%), DM(4.2%). Accompanied extrapulmonary tuberculosis were pleurisy(5.4% ), superficial lymphadenitis(0.4% ). CONCLUSIONS: There was great effects in the treatment of tuberculosis with short course regimen in the National Tuberculosis Control Program. But only 38% among the expected patients were treated in this country. So the greater efforts were needed to find and treat more patients effectively.

A Case Report of Double Outlet Right Ventricle(S.D.L.) with Subpulmonic Ventricular Septal Defect and Pulmonary Stenosis

No abstract available.

An experimental study on the alteration of thermal enhancement ratio by combination of split dose hyperthermia irradiation

The study was undertaken to evaluate the alteration of thermal enhancement ratio as a function of timeintervals between two split dose hyperthermias followed by irradiation. For the experiments, 330 mice were dividedinto 3 groups; the first, 72 mice were used to evaluate the heat reaction by single dose hyperthermia and heatresistance by split dose hyperthermia, the second, 36 mice were used to evaluate the radiation reaction byirradiation only, and the third, 222 mice were used for TER observation by comibination of single dosehyperthermia and irradiation , and TER alteration by combination of split dose hyperthermia and irradiation. Foreach group the skin reaction score of mouse tail was used for observation and evaluation of the results of heatand irradiation. The results obtained are summarized as follows; 1. The heating time resulting 50% necrosis(ND 50)was 101 minutes in 43degrees centigrade and 24 minutes in 45degrees centigrade hyperthermia, which indicated thatthere is reciprocal proportion between temperature and heating time. 2. Development of heat resistance wasobserved by split dose hyperthermia. 3. The degree of skin reaction by irradiation only was increasedporportionally as a function of radiation dose, and calculated radiation dose corresponding to skin score 1.5(D1.5) was 4,137 rads. 4. Obtained thermal enhancement ratio by combination of single dose hyperthermia andirradiation was increased proportionally as a function of heating time. 5. Thermal enhancement ratio was decreasedby combination of split dose hyperthermia and irradiation , which was less intense and lasted longer thandevelopment of heat resistance. In summary, these studies indicate that the alteration of thermal enhancementratio has influence on heat resistance by split dose hyperthermia and irradiation.

Radiologic study of silicosis in Korean

These radiologic studies were carried out on 265 cases of silicosis which were diagnosed clinically atindustrial Accident Hospital affiliated with Catholic Medical Collage, suring the period of 3 years from 1974 to1976. 265 cases of silicosis consisted of 96 cases (35%) of coal miner, 91 cases (34%) of coal choicer and 82cases (31%) of others. The average age was 42.2 years and average working period was 9.2years. Qualitative andquantitative features in the analysis of roentgen findings were based on UICC/Cincinnati and KLO classification.The qualitative features showed 26% of "p" opacity, 46% of "q" opacity 4% of "r" opacity as rounded profusion and5% of "s" opacity, 11% of "t" opacity, 2% of "u" opacity as irregular profusion of small opacities.Large opacitiesshowed 3% of group A, 2% of group B and 1% of group C. Quantitative features revealed 25% of Type 1, 54% of Type2, 16% of Type 3 and 5% of Type 4. The qualitative and quantitative features showed significant differences as ageand working period increased with age and working period. Major complications were pulmonary emphysema,tuberculosis and pleural abnormalities.

Three Cases of Acneiform Demodicidosis due to Long - Term Application of Corticosteroid Ointment / 대한피부과학회지

Demodicidosis is caused by the enormous infestation of 400 micron sized demodex folliculorum which normally exist, though in small number, in pilosebaceous unit. Its characteristic clinical feature is erythema and erythematous indurated papulopustular eruptions combined with itching and buming sensation. Microscopic examination of indurated pustule always reveals demodex folliculorum. Many authors have reported that demodicidosis aggravated by avoidance of using soap, long-term use of cosmetic cream, especially cold cream, topical or systemic use of corticosteroid. Our patiednts were housewives of 46,39, and 42 years old and presented characteristic clinical featurss and skin lesions by topical application of corsicosteroid for 1~2 years and were diagnosed as steroid acne. But simple microscopic examination reveled demodex folliculorum. We treated them with 5% sulfur lintment and soap and water cleansing of face with remarkable clinical improvement.

Clinical Effect of Glutathione (Tathion) on Melasma / 대한피부과학회지

Tatbion is a tripeptide, reduced form of Glutathione(GSH or p-glutamyl-cysteiny1 -glycine). Glutathione(SH compound)is believed to inhibit melanin formation by combining witb the copper in tyrnsinase whicb is essential in the conversion of tyrosine to DOPA (3, 4-dihydroxyphenylalanine) and DOPA to DOPA-quinone or by forming cornplex with the intermediate in the tyrosine-to-melaa.in reaction. The effect of Tathion in the treatment of melasma has not been reported in Korea. We have observed the effect of Tatbion in 150 patients with melasma. After the average duration of Gwks of treatment(50-100mg tree tirnes/daily), we were abIe to grade the results as follow. Excellent(Pigmentation almost disappeared): 17. 4 % Good (Pigmentation markedly improved): 56. 7% Fair (Pigmentation slightly improved: 7. 3% None (No effect) : 18.6% The result showed relatively good effects of Tathion in the treatment of 122pts (81.4%) with melasma in total. The brief review of literature on the treatment of melasma was undertaken.

Clinical Observation of Epidermolysis Bullosa Dystrophica / 대한피부과학회지

Epidermolysis bullosa Dystrophica is rare, chronic non-mflammatory mechanobullous disease of hereditary trait, which easily produces bulIa by minor trauma or spontaneousIy. The lesions usually appears from birth or in infant involving especially lower leg, hand, foot and lea,ds to erosive, hemorrhagic bulla, secondary infection, pruritus and scar formation and sometimes deformity of nails and cicatrical alopecia. It sometimes involves oral mucosa, esophagus, eye, tooth, bone, respiratory system, anus and vagina. This disease has rare tendency for carcinogenesis from cicatrical Iesions. These manifestations are more severe in recessive type than in dominant type. We had observed 5 cases of EBD at Ewha Womans University Hospital from Sept. 1975. to Jul. 1976. Among 5 cases, 4 cases occurred from birth, 1 case occurred at age l. Skin lesivns were hand, foot, knee area in all 5 cases. Head, face were involved in 2 cases. Arm, leg, trunk, buttock were involved in 3 cases. In skin manifestations, bulla, erosive bulla, secondary infection, scar formation, pruritus, nail deformity, positive Nikolsky sign were observed in all 5 cases. Hemorrhagic bulla, miliaria. were presented, in 1 case, alopecia in 2 cases. Aggrevated season were summer in 4 cases, spring in 1 case, HistologicalIy, microscopic observation, after H@-E stain, revealed hyperkeratosis in 3 cases, acanthosis in 2 cases, partial atrophy in 1 case, partial ulcer in 1 case, snbepidermal bulla in 5 cases, chronic nonspecific inflamma- tory cell infiltration of upper dermis in 3 cases, fibrosis of lower dermis in 1 case.

Statistical Consideration in Syphilitic Patients in Ewha Womans University Hospital (1974. 8. - 1977. 8.) / 대한피부과학회지

The recent prevalence of syphilis has known to be increased progressively throughout the world, since middle of 1950. In Korea, current trend of increased syphilic infection has became a major public health problem. The authors selected sero-positive patients, among the outpaients of department of dermatology, urology, OB & GY, and physical examination, and performed statistical analysis of this sero-positive patients. Results are as following; 1) Of the 2,007 patients taken physical examination, 30 patient(1.49%) were reactive to VDRL. 2) 0.71% of the total out patients of the dermatologic dcpartment were sero-positive. Annual ratio of the syphilitic patient were increased progressively. 3) Ratio of the syphilitic patients to the total outpatients of OB & GY department was 0.55% and somewhat increased since 1976. 4) In the age distribution, the third decade shared 51.8% of the total sero-positive patients 5) In the sexual ratio. male:female=l: l. 62 in the total sero-positive patients. (male:female=l: 1.06 in the dermatologic department.) 6) In the stage distribution, latent syphilis patients were 59.14% of the total syphilitic patients.

A Case of Juvenile Xanthogranuloma / 대한피부과학회지

Juvenile Xan anuloma is granulomatous, benign disseminated Xanthomatous disease, which i r cytosis of unknown origin, due to lipid, metabolism disturbance. Spontaneous remi. is possible authors have reported some cases of JXG involving only skin or combii extracutaneous lesions, In our country, cases involving onIy skin have bcen reported. This I-year-old norered healthy male baby has yellowish brown coloxed pinhead to rice sizeci gers of well defined. round, dome shaped smooth surface in face, trunk, lower abdomen without any subjective symptomes. Serum lipid level i iid chest p-A,urinalysis, CBC, L.F.T. are normal. Family history showed ific finding and physical examina.tion shows nothing remarkable except for skeen lesions. EIistopathologically, hi;tiocyte, lymphocyte, eosinophile, foam cell, foreign body giant cell, typical Toutor giant cell are seen.

A Case of Verrucous Hemangioma / 대한피부과학회지

Verrucous hemangioma is congenital hemangioma, which is structural varients of capillary or cavernous hemangioma. And it is vascular malformation, in which reactive epidermal acanthosis, papillomatosis, hyperkeratosis develope secondarily. Though foreign anthors have reported some cases of this disease, it is rare hemangioma and it never has been reported in Korea. This 13 year-old, healthy female patient has erythematous patchs on her left knee at birth. It grew become coin to egg sized. Because of verrcous iuflammatory reaction of irregular verrucous surfaced plague, the lesion become necrotic thick crusty plague with severe offensive odor and patient complained of severe pain & motion limitation on her left knee. Urinalysis, C B C, chest PA were within normal limit. PPD test was negative. Lt. knee AP & lat. revealed. Marked destroyed soft tissue on anterior portion of Lt. knee and no bony pathological changes were demonstrable. Histopathologica.11y, hyperkeratosis, parakeratosis, acanthosis, papillomatosis in the epidermis and numerous capillary lumina, capillary dilatation, proliferation of endothelial cells and mild infiltration of inflammatory cells in the Dermis. And fibrosis in the Dermis & Subcutaneous tissue are seen.

A Case of Darier's Disease / 대한피부과학회지

Darier's diseaae is relatively rare dyskeratotic and inheritable dermatoses of an. autosomal irregular dominant gene. The authors observed one case of typical Dariers disease with the family hietory of three generatian of autoaomal daminant trait. 22 year-oid patient we observed has had the generalized pruritus and dark dirty and warty patehes distribnted almost on the whole skin surface especially on trunk, back and face for about 10 years. Histopathologically, the lesion showed lacunae, corps ronds and grains which are compatible with typical Dariers disease.