Ryou Misao

Revised Guidelines for Neonatal Screening Programmes for Primary Congenital Hypothyroidism

Since the first guidelines for neonatal screening for congenital hypothyroidism (CH) were issued by ESPE in 1993 [1], there have been considerable advances in our understanding of CH and our appreciation of the various geographical and logistic difficulties involved. Therefore, an updating of the guidelines is overdue. Experience from countries where screening began in the late 1970s and early 1980s has indicated that treatment should be started no later than the first 2 weeks of life using a ‘high’ dosage regime of L-thyroxine (10–15 Ìg/kg/day). It has also been shown that the quality of long-term outcome is closely related to the quality of follow-up. In Eastern Europe, screening programmes for CH have either been started or will start soon in almost all countries, and although many programmes are operating satisfactorily, it is important to standardise screening procedures and management of suspected cases as much as possible in order to optimise outcome. A degree of uniformity throughout Europe would not only facilitate early detection and treatment of individual patients but give insight into the economic and epidemiological aspects of the screening programmes as well as the epidemiological aspect of CH.

Dominant expression of progesterone receptor form B mRNA in ovarian endometriosis.

This study was designed to examine the biological implications of progesterone receptor form A (PR-A) and B (PR-B) mRNA expressions in human ovarian endometriosis (ectopic endometrium). A high ratio of PR-B to PR-AB (PR-A+PR-B) mRNA expression was found in 8 of 14 cases of endometriosis, compared with the ratio in eutopic endometrium. The mean ratio in ectopic endometria was significantly (p < 0.01) higher than in eutopic endometria. The ratio in eutopic and ectopic endometria showed no significant change during the menstrual cycle. The mean ratio in ectopic endometria in the proliferative and secretory phases of the endometrium was significantly (p < 0.01) higher than in eutopic endometria. In conclusion, PR-B mRNA was relatively highly expressed in some endometriomas, which might lead to aberrations in the control of progestational effects involving responsiveness to sex steroidal growth regulation.

Evidence for the Synthesis of Corticosteroid-Binding Globulin in Human Placenta

We demonstrated the expression of corticosteroid-binding globulin (CBG) in human placenta using reverse transcription-polymerase chain reaction-Southern blot analysis and immunohistochemical and immunoblotting studies. In the RT-PCR-Southern blot analysis, only one predicted PCR product was detected without nonspecific products in all samples of human placenta and 3A (tPA-30-1) human placental cells. In Western blot analysis, polyclonal anti-CBG antibodies recognized a protein of approximately 55 kD in the protein extracts prepared from 3A (tPA-30-1) cells. Additionally, CBG mRNA expression was demonstrated by in situ hybridization in the syncytiotrophoblasts. Immunohistochemical studies performed on the placenta demonstrated the presence of specific immunoreactivity in the syncytiotrophoblast layer surrounding the chorionic villi. These findings suggest that CBG is synthesized in human placenta during pregnancy in addition to its synthesis in the liver.

Identification of exon-deleted progesterone receptor mRNAs in human uterine endometrial cancers

We demonstrated the expression of various exon-deleted progesterone receptor (PR) variant mRNAs in human uterine endometrial cancers using the reverse transcription-polymerase chain reaction-DNA sequencing analyses. In addition to PR wild-type mRNA, exon 4-deleted, exon 6-deleted, exon 3,4-deleted, exon 5,6-deleted, exon 4,5,6-deleted and exon 3,4,5,6-deleted PR variant mRNAs were identified. The exon 6-deleted and exon 5,6-deleted PR variant mRNAs lacked encoding for the steroid-binding domain. The exon 4-deleted, exon 3,4-deleted, exon 4,5,6-deleted and exon 3,4,5,6-deleted PR variant mRNAs lacked encoding for the DNA-binding domain in addition to encoding for the steroid-binding domain. While the exon 4-deleted, exon 6-deleted and exon 3,4-deleted PR variant mRNAs were observed in all samples analyzed, the exon 5,6-deleted, exon 4,5,6-deleted and/or exon 3,4,5,6-deleted PR variant mRNAs could not be detected in some cases, especially in poorly differentiated adenocarcinoma as compared with well-differentiated and moderately differentiated adenocarcinomas. The present study demonstrates the coexpression of PR exon-deleted variant mRNAs with the wild-type in uterine endometrial cancers. All translated variant proteins might possess functional diversity and might modify the progestational action of wild-type PR, and the expression of some PR variant mRNAs may be lost as endometrial cancer cells undergo dedifferentiation.

Corticosteroid-binding globulin mRNA levels in human uterine endometrium

Corticosteroid-binding globulin (CBG or transcortin) is a specific plasma glycoprotein, which binds steroid hormones (cortisol, corticosterone, and progesterone), and plays a role in transporting these steroids, altering their concentrations in blood, and influencing their biological actions. CBG has been previously shown to be synthesized in the liver, but recently it has been reported that immunoreactive CBG is localized in target tissues. In the present work, CBG mRNA was detected in normal human endometrial tissues by Northern blot analysis and reverse transcription-polymerase chain reaction. Its level was higher (P < 0.05) in the secretory phase than in the proliferative phase. In the secretory phase, the endometrial CBG mRNA level was negatively correlated with the serum progesterone level (P < 0.01). While there was no positive correlation between the levels of endometrial CBG mRNA and serum estradiol (E2), there was a positive correlation between the endometrial CBG mRNA level and the serum E2/progesterone ratio (P < 0.05). These findings suggest that CBG is synthesized in the uterine endometrium, predominantly in the secretory phase, and that the serum E2/progesterone ratio exerts an influence on the synthesis of intracellular CBG.

Expression of sex hormone-binding globulin mRNA in human endometrial cancers

To more fully understand the role of sex hormone-binding globulin (SHBG) on the intracellular steroidal action in endometrial cancers, we investigated the expression of SHBG mRNA as the substitute of SHBG expression in human endometrial cancers. In the present study, the levels of SHBG mRNA were analyzed using competitive reverse transcription-polymerase chain reaction (RT-PCR)-Southern-blot analysis. The higher level of SHBG mRNA tended to be expressed in the normal secretory and late proliferative phase endometrium > early proliferative phase endometrium > well differentiated adenocarcinoma of the endometrium (G1) > moderately differentiated adenocarcinoma (G2) > poorly differentiated adenocarcinoma (G3), in the order shown. These studies indicate that endometrial cancer cells might synthesize intracellular SHBG to conserve their estrogen-dependent properties. Further, it indicates that endometrial cancer cell synthesis of SHBG mRNA is lost as these cells undergo de-differentiation.

Expression of sex hormone-binding globulin exon VII splicing variant mRNA in human uterine endometrium

We have demonstrated the expression of sex hormone-binding globulin (SHBG) exon VII splicing variant mRNA in human uterine endometrium, using the reverse transcription-polymerase chain reaction-Southern blot and DNA sequencing analyses. Analysis of the missing base pairs corresponded to the entire exon VII, which are considered to encode a portion of the steroid-binding site. Therefore, the steroid-binding affinity of this variant might be different from that of the SHBG wild type. In uterine endometria, the wild-type and variant mRNA levels tended to increase with the advance of the menstrual phase, but the ratio of the SHBG variant mRNA to SHBG wild-type mRNA levels showed no significant difference during the menstrual cycle. So far, there are no indications that the SHBG variant has any biological or clinical implications in human uterine endometrium.

Steroid receptor mRNA levels in human corpus luteum.

To understand the biology of sex steroids in the human corpus luteum, the expression of estrogen receptor alpha, progesterone receptor, and androgen receptor mRNA levels was determined by semiquantitative reverse-transcription polymerase chain reaction-Southern blot analysis. Expression of all receptor mRNAs was detected in all samples analyzed. Each steroid receptor mRNA level was significantly lower (p < 0.05) during the late secretory phase than that during the early or the mid-secretory phase of the endometrium. These findings support the concept of a local role for sex steroids in modulating the function and life span of the human corpus luteum.

Expression of Sex Hormone-Binding Globulin and Corticosteroid-Binding Globulin mRNAs in Corpus luteum of Human Subjects

To understand the biology of sex steroids in human ovarian corpus luteum, the expression of intracellular sex hormone-binding globulin (SHBG) and corticosteroid-binding globulin (CBG) mRNAs as a manifestation of intracellular SHBG and CBG expression was determined. The expression of SHBG and CBG mRNAs was detected in all samples analyzed. Luteal SHBG mRNA level showed no significant change during the endometrial secretory phase of the menstrual cycle. On the other hand, luteal CBG mRNA level was significantly higher (p < 0.05) at the mid-secretory phase than that at the early and late secretory phases of the endometrium. These findings suggest that human ovarian corpus luteum synthesizes SHBG and CBG intracellularly, CBG being plausibly involved in the functional life span of corpus luteum.

Expression of sex hormone-binding globulin exon VII splicing variant messenger ribonucleic acid in human ovarian endometriosis

OBJECTIVE To investigate the expression of sex hormone-binding globulin (SHBG) exon VII splicing variant messenger RNA (mRNA) in human ovarian endometriosis. DESIGN The expression of SHBG variant mRNA in normal uterine endometrium and endometriotic tissue was determined. SETTING Department of Obstetrics and Gynecology, Gifu University Hospital. PATIENT(S) Fourteen women with endometriosis and 18 women without endometriosis. INTERVENTION(S) Normal uterine endometrial and ovarian endometriotic tissues from patients who had undergone gynecological surgery were studied. MAIN OUTCOME MEASURE(S) Levels of SHBG wild-type and variant mRNAs were determined using the quantitative reverse transcription-polymerase chain reaction. RESULTS(S) Analysis of the missing base pairs proved that they corresponded to the entire exon VII. There was no significant difference between the levels of SHBG wild-type mRNA in normal endometria and in endometriotic endometria, although the levels of SHBG variant mRNA in endometriotic endometria were significantly higher than that in normal endometria. The ratio of SHBG variant to wild-type mRNA levels was significantly higher in endometriotic endometria than in normal endometria. CONCLUSION(S) This study demonstrates the coexpression of SHBG exon VII splicing variant mRNA with its wild-type and the dominant expression of the variant in ovarian endometriosis. These results might be involved in the cellular estrogenic interaction, plausibly assisting in the development and growth of ovarian endometriosis.