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The objective of this study was to use fluorescence-activated cell sorting (FACS) and spermatogonial stem cell (SSC) xenotransplantation to identify cell surface markers of putative porcine SSC. Analysis of porcine testis cells enriched for spermatogonia using FACS indicated that nearly half of stage-specific embryonic antigen-1 (SSEA-1) expressing testis cells expressed the undifferentiated spermatogonia marker protein gene product 9.5 (PGP 9.5) whereas significantly fewer (P < 0.05) cells selected for thymus cell antigen-1 (Thy-1), also known as cluster of differentiation 90 (CD90), cluster of differentiation 9 (CD9), or other SSC markers expressed PGP 9.5. Immunocytochemical analysis indicated that promyelocytic leukemia zinc finger (PLZF) protein and germ cell lineage marker VASA homolog (VASA), also known as DEAD box protein 4 (DDX4), were expressed by SSEA-1 expressing germ cells. Spermatogonial stem cell xenotransplantation of testis cell populations enriched for cells expressing SSEA-1 generated significantly (P < 0.05; greater than 15-fold) more colonies of donor derived germ cells than unselected testis cells. In conclusion, these data indicate that SSC markers identified in rodents are likely not entirely conserved in pigs and that SSEA-1 is a marker for porcine undifferentiated spermatogonia including SSC in prepubertal boars and its expression may serve as a target for the further study of porcine germ cells.

Adipose triglyceride lipase (ATGL), a newly identified lipase, is a rate-limiting enzyme for triglyceride hydrolysis in adipocytes. The regula- tory proteins involved in ATGL-mediated lipolysis in fat tissue are not fully identified and understood. The G(0)/G(1) switch gene 2 (G0S2) is an inhibitor of ATGL activity by interacting with ATGL through the hydrophobic domain of G0S2. Here, for the first time, we have cloned the coding sequence of G0S2 cDNA for the chicken, turkey, and quail. Sequence compari- sons with mammals revealed that the avian G0S2 also have a conserved hydrophobic domain. Avian G0S2 is predominantly expressed in adipose tissues relative to other tested tissues. Within the adipose tissue, G0S2 is expressed 20-fold greater in the adipocyte than in the stromal-vascular (SV) fraction (P < 0.001). Expression of G0S2 mRNA gradually increased during differentia- tion of chicken adipocytes in culture (P < 0.05). How- ever, there is G0S2 expression in embryonic adipose tissue, SV fraction, and primary preadipocytes before confluence that generally have an increased capacity of cell proliferation, which indicates it has an important role in adipocyte differentiation rather than prolifera- tion. For a better understanding of how G0S2 responds to environmental stimuli, chickens were fasted for 24 h and then refed. Expression of G0S2 in adipose tissue was dramatically decreased (P < 0.05) in the chickens and quail after a 24-h fasting period, and increased to the control level after refeeding. In contrast to G0S2 ex- pression, ATGL expression was induced (P < 0.05) af- ter the 24-h fasting period and rapidly returned to the control level during the refeeding period. These data indicate that changes in lipolytic activities of adipose tissue in vivo can be regulated by G0S2 expression, as an inhibitor of ATGL.

The objective of this study was to use fluorescence-activated cell sorting (FACS) and spermatogonial stem cell (SSC) xenotransplantation to identify cell surface markers of putative porcine SSC. Analysis of porcine testis cells enriched for spermatogonia using FACS indicated that nearly half of stage-specific embryonic antigen-1 (SSEA-1) expressing testis cells expressed the undifferentiated spermatogonia marker protein gene product 9.5 (PGP 9.5) whereas significantly fewer (P0.05) cells selected for thymus cell antigen-1 (Thy-1), also known as cluster of differentiation 90 (CD90), cluster of differentiation 9 (CD9), or other SSC markers expressed PGP 9.5. Immunocytochemical analysis indicated that promyelocytic leukemia zinc finger (PLZF) protein and germ cell lineage marker VASA homolog (VASA), also known as DEAD box protein 4 (DDX4), were expressed by SSEA-1 expressing germ cells. Spermatogonial stem cell xenotransplantation of testis cell populations enriched for cells expressing SSEA-1 generated significantly (P0.05; greater than 15-fold) more colonies of donor derived germ cells than unselected testis cells. In conclusion, these data indicate that SSC markers identified in rodents are likely not entirely conserved in pigs and that SSEA-1 is a marker for porcine undifferentiated spermatogonia including SSC in prepubertal boars and its expression may serve as a target for the further study of porcine germ cells.

Adipose triglyceride lipase (ATGL), a newly identified lipase, is a rate-limiting enzyme for triglyceride hydrolysis in adipocytes. The regulatory proteins involved in ATGL-mediated lipolysis in fat tissue are not fully identified and understood. The G(0)/G(1) switch gene 2 (G0S2) is an inhibitor of ATGL activity by interacting with ATGL through the hydrophobic domain of G0S2. Here, for the first time, we have cloned the coding sequence of G0S2 cDNA for the chicken, turkey, and quail. Sequence comparisons with mammals revealed that the avian G0S2 also have a conserved hydrophobic domain. Avian G0S2 is predominantly expressed in adipose tissues relative to other tested tissues. Within the adipose tissue, G0S2 is expressed 20-fold greater in the adipocyte than in the stromal-vascular (SV) fraction (P0.001). Expression of G0S2 mRNA gradually increased during differentiation of chicken adipocytes in culture (P0.05). However, there is G0S2 expression in embryonic adipose tissue, SV fraction, and primary preadipocytes before confluence that generally have an increased capacity of cell proliferation, which indicates it has an important role in adipocyte differentiation rather than proliferation. For a better understanding of how G0S2 responds to environmental stimuli, chickens were fasted for 24 h and then refed. Expression of G0S2 in adipose tissue was dramatically decreased (P0.05) in the chickens and quail after a 24-h fasting period, and increased to the control level after refeeding. In contrast to G0S2 expression, ATGL expression was induced (P0.05) after the 24-h fasting period and rapidly returned to the control level during the refeeding period. These data indicate that changes in lipolytic activities of adipose tissue in vivo can be regulated by G0S2 expression, as an inhibitor of ATGL.

To investigate the high circumcision rate in South Korea and its rapid increase in the short period since its introduction.From January to December 2000, 5434 South Korean males (or their parents) aged 0-92 years were interviewed in detail about their circumcision status, their age at circumcision, and the possible effect of circumcision on their sexuality. In addition, 267 practising medical doctors were surveyed about their basic understanding of circumcision and phimosis.Currently the circumcision rate for high-school boys is90% and for those70 years old is10%. The circumcision rate in 1945 was0.1%. When averaged over the whole population, the present South Korean circumcision rate is approximately 60%; the rate has increased dramatically with time and particularly in the past 20 years, when the estimated number of male circumcisions has exceeded the number of male births. Although circumcision in South Korea has been strongly influenced by American culture, it has never been predominantly neonatal. The age at circumcision has continued to decrease and boys are now circumcised at approximately 12 years old. Of those who were circumcised long after they had been sexually active,80% reported no noticeable difference in sexuality, but a man was twice as likely to have experienced diminished sexuality than improved sexuality. Of the doctors who were surveyed, 41% carried out circumcision but, unlike in America, gynaecologists and paediatricians rarely did so. Among the doctors, basic knowledge on circumcision and phimosis was generally lacking, regardless of whether they practised circumcision or not. Amongst the factors contributing to the high circumcision rate was the mistaken notion held by both doctors and the general public that circumcision is directly correlated with industrialization and general progress of living standards. Many doctors believe the out-dated and sometimes controversial benefits of circumcision, i.e. prevention of cervical cancer and sexually transmitted diseases, and improved sexuality. Thus the vast majority of doctors recommend circumcision regardless of the patient's age. Peer pressure was also an important contributing factor.South Korea has an unusual history of circumcision. The mistaken and out-dated notions about circumcision and lack of knowledge of phimosis by physicians seem to be a leading contributory factor to the extraordinarily high circumcision rate.

We have developed a transgenic female goat harboring goat beta-casein promoter/human granulocyte colony stimulating factor (G-CSF) fusion gene by microinjection into fertilized one-cell goat zygotes. Human G-CSF was produced at levels of up to 50 microg/ml in transgenic goat milk. Its biological activity was equivalent to recombinant human G-CSF expressed from Chinese hamster ovary (CHO) cell when assayed using in vitro HL-60 cell proliferation. Human G-CSF from transgenic goat milk increased the total number of white blood cells in C57BL/6N mice with leucopenia induced by cyclophosphamide (CPA). The secreted human G-CSF was glycosylated although the degree of O-glycosylation was lower compared to CHO cell-derived human G-CSF.