Search

BACKGROUND: Klinefelter's syndrome is the most frequent chromosomal abnormality in infertile men. In this study, the chromosomes of round spermatids and spermatogonia/primary spermatocytes from men with non-mosaic Klinefelter's syndrome were examined, together with the Sertoli cell secretory function and sperm morphometry. METHODS: Twenty-four men with non-mosaic Klinefelter's syndrome and nine men with obstructive azoospermia underwent therapeutic testicular biopsy. When spermatozoa in the final filtrate were present, they were processed for sperm morphometry or ICSI. Sperm morphometry was evaluated by the maximal length and width of the sperm head, the length of the midpiece and the ratio of the acrosomal region to the total surface area of the head. When round spermatids were present, they were processed for fluorescent in-situ hybridization (FISH). FISH was also applied to fragments of seminiferous tubules. Sertoli cell secretory function was measured by the amount of androgen binding protein (ABP) secreted in vitro. RESULTS: More than 93% of the evaluated round spermatids were normal. The proportions of 24,XY and of 24,XX round spermatids to the total number were significantly larger in men with Klinefelter's syndrome than in obstructed azoospermic men. Men with Klinefelter's syndrome who had spermatozoa in their testicular tissue (n = 12) were positive for both 46,XY and 47,XXY spermatogonia in their seminiferous tubules. In contrast, men with Klinefelter's syndrome without spermatozoa in their testicular tissue (n = 12) were positive for 47,XXY spermatogonia but negative for 46,XY spermatogonia in their seminiferous tubules. ABP profiles were significantly smaller in men with Klinefelter's syndrome who were negative for spermatozoa compared with men who were positive. Four pregnancies were achieved and five healthy babies were born. CONCLUSIONS: This study suggests that few 46,XXY spermatogonia undergo meiosis with an XX pairing and a Y univalent type of pairing. Hyperhaploid round spermatids (24,XY and 24,XX) may be produced by meiosis of 47,XXY spermatogonia. Men with Klinefelter's syndrome who are negative for testicular spermatozoa have a greater degree of Sertoli cell secretory dysfunction compared with men with Klinefelter's syndrome who are positive for spermatozoa. There are several defects in sperm morphometry with functional significance in men with Klinefelter's syndrome.

Klinefelter's syndrome is the most frequent chromosomal abnormality in infertile men. In this study, the chromosomes of round spermatids and spermatogonia/primary spermatocytes from men with non-mosaic Klinefelter's syndrome were examined, together with the Sertoli cell secretory function and sperm morphometry.

The primary nucleation rate I and radial growth rate G of the isothermal spherulitic growth of poly(ethylene oxide) in tripropionin solutions of 5, 10, and 30 wt% polymer concentrations were studied by direct in situ observation using a CCD camera. I was found to decrease with pre-annealing at 200°C up to ca. 200 h, while G was almost constant. These results suggest that there exist micro-residuals of a crystalline-like structure that act as dominant seeds for primary nucleation and that they become dissolved with structural relaxation during pre-annealing, resulting in the solution approaching a homogeneous state. The supercooling dependencies of I and G were independently investigated, and it was found that the fold surface free energy and activation energy of the polymer diffusion increase with decreases in the polymer content of the solution. The frictional interaction between the polymer and solvent is also discussed on the basis of the reaction rate theory of Kramers.

For clinical islet cell transplantation, short-term storage of islet cells is likely to be necessary, and it is imperative that the islet cells be kept as viable as possible during the period. However, there are little data on which preservative solutions are most suitable for the storage of islet cells after isolations or before transplantation. To estimate islet cell viability and transplantation success rate in the present study, adenylylcyclase activity was measured with a rapid new fluorometric assay in rat islet cells prior to transplantation, because cAMP plays an essential role in determining islet β-cell viability and responsiveness to various hormonal stimuli. Adenylylcyclase activity was measured in islet cells stored for different periods of time (0, 3, 16, 24, 48, 96 h) and in different preservative solutions. Approximately 1,000 islet cells from each preservation group using University of Wisconsin (UW) solution were transplanted to streptozotocin-induced diabetes mellitus (DM) rats. Transplant success was evaluated by measuring blood glucose levels. Preoperative adenylylcyclase activity was compared with posttransplant islet cell function. The adenylylcyclase activity of UW solution was significantly higher than that of Euro-Collins solution and lactate-Ringer's solution through the different preservation time periods. Preoperative adenylylcyclase activity correlated well with posttransplant islet cell function in a rat model of DM. We conclude that adenylylcyclase activity can be used as a marker to assess islet cell viability as well as differences in preservation media and may predict islet cell transplant success.

The effects of hypercholesterolaemia on testicular endocrine and exocrine function were evaluated. The influence of hypercholesterolaemia on sperm quality, quantity, and fertilizing potential was also determined. Ten mature rabbits (group A) were fed chow containing 3% cholesterol for 12 weeks. Ten control rabbits (group B) were fed normal chow for the same period. At the end of the experimental period testosterone profiles and sperm parameters were evaluated and the sperm reproductive potential was assessed by in vitro fertilization (IVF) techniques. Peripheral serum testosterone responses to testicular stimulation with human chorionic gonadotrophin, androgen-binding protein activity in testicular cytosols, sperm concentration, sperm motility, length of sperm midpiece, and IVF outcome were all significantly lower in group A than in group B. In contrast, serum cholesterol concentrations were significantly higher in group A. There were no significant differences in either testicular versus intra-abdominal temperature differences or cholesterol concentrations in seminal plasma or testicular tissue between groups A and B. The results suggest that hypercholesterolaemia has a detrimental effect on Leydig and Sertoli cell secretory function, spermatogenesis, epididymal sperm maturation process, and the overall sperm fertilizing capacity.

We compared the outcome of ooplasmic round spermatid nuclear injections (ROSNI) versus intact round spermatid injections (ROSI). Rabbit round spermatid nuclei and intact round spermatids were recovered and injected into rabbit oocytes (groups A and B, respectively). Fertilization, cleavage and embryonic development rates were compared. In additional studies, five protocols for cryopreservation of round spermatids and two protocols for cryostorage of round spermatids were applied. The outcome of ROSNI techniques using frozen–thawed or cryostored–warmed round spermatids was evaluated. The cleavage rate and the overall morula plus blastocyst development rate were significantly larger in group A than group B. ROSNI procedures are superior to ROSI techniques in the rabbit. The largest fertilization, cleavage and embryonic development rates after ROSNI techniques using cryopreserved or cryostored round spermatids were demonstrated in groups of round spermatids in which a mixture of seminal plasma plus test yolk buffer was employed as an extender, and dimethyl sulphoxide plus a high concentration of glycerol served as cryoprotectants. It appears that the seminal plasma contains factors protecting round spermatids during cryopreservation or cryostorage, and/or the employment of two cryoprotectants has a beneficial role in the maintenance of round spermatid reproductive capacity.

The role of a telomerase assay in the recognition of Sertoli cell-only syndrome with testicular foci of haploid cells was evaluated. Men with Sertoli cell-only syndrome (n = 23) were given a new diagnostic testicular biopsy. Part of the biopsy was stained and the remainder was processed for the quantitative telomerase assay. After 3–13 months, a therapeutic testicular biopsy was performed. This material was minced and then examined using confocal laser scanning microscopy and fluorescent in-situ hybridization. Histology of diagnostic testicular biopsy material confirmed the diagnosis of Sertoli cell-only syndrome in all the participants. All seven men with a telomerase assay value in their diagnostic testicular biopsy of >42 total product generated (TPG) U/μg protein had haploid cells (i.e. spermatozoa and/or spermatids) in their therapeutic testicular biopsy. Among participants with telomerase assay values <42 TPG U/μg protein, only one man had haploid cells in his therapeutic testicular biopsy. Thus, telomerase assay values >42 TPG U/μg protein in the diagnostic biopsy identified 87.5% of the Sertoli cell-only syndrome men with haploid cells in their therapeutic testicular biopsy. Significantly higher values of the telomerase assay were found in men with testicular foci of haploid cells than in men without these foci. The use of a quantitative telomerase assay biopsy appears to be important for identifying those men with Sertoli cell-only syndrome who have foci of haploid cells and can be candidates for assisted reproduction techniques.

Abstract: We investigated whether testicular telomerase activity is due to telomerase expression in all cells or expression in a limited number of cells. Telomerase activity was assayed in highly purified fractions of spermatogonia cells plus primary spermatocytes, secondary spermatocytes plus round spermatids, secondary spermatocytes plus spermatids plus spermatozoa, round spermatids, or spermatozoa prepared from healthy or cryptorchid animals. Telomerase activity was additionally assayed in testicular tissue of prepubertal animals and animals with Sertoli cell only pathophysiology. Telomerase activity was detected in fractions containing primary spermatocytes and/or secondary spermatocytes and/or spermatids. Fractions enriched in round spermatids were positive for telomerase activity. In contrast, spermatozoa or Sertoli cell fractions were negative for telomerase activity. Using the relative telomerase activity assay and the sensitive quantitative telomerase assay to quantify telomerase activity, we showed that induction of cryptorchidism does not result in quantitative alterations in testicular tissue telomerase activity. In addition, elimination of round spermatids does not lead to significant alterations in testicular tissue telomerase activity. The present results suggest that the male gamete telomerase activity is inhibited during spermiogenesis. Furthermore, it appears that spermatogonia/primary spermatocytes are the main sources of telomerase activity in the testis.

Spherulitic growth of poly(ethylene oxide) from 30 wt% tripropionin (glycerol tri-n-propionate) solution was investigated in the supercooling range ?T=11–37 K. The solution was pre-annealed at 200°C before isothermal crystallization. The estimated total crystallization rate decreases with increasing pre-annealing time, and becomes almost constant with a pre-annealing time longer than ca. 200 h. This suggests that traces of crystalline nucleus remaining in the solution dissolve during the pre-annealing process. The isothermal crystallization rate G with respect to the crystallization temperature was measured using isothermal calorimetric measurements and direct observation of the spherulitic growth using a CCD camera. The Avrami exponent estimated from the isothermal crystallization data ranges from ca. 2.7 to 3.2 depending on the crystallization temperature. The profile of G vs. T was also analyzed to estimate the parameters for the diffusion factor, and the result shows that the diffusion process belongs to the low friction limit, i.e., frictional interaction between the polymer and solvent is extremely weak.

Abstract:: To determine the tumor size that constitutes early pancreatic cancer, we reviewed and analyzed the English-language and Japanese literature (a total of 25 publications) on small pancreatic cancers less than 2 cm in diameter and/or stage 1 cancers. Reports on in situ carcinoma and intraductal carcinoma of the pancreas were also evaluated. The results were: (1) A total of 302 cases of small pancreatic cancer less than 2 cm in diameter reported at separate institutions were pooled from 15 reports. The rates for patients in stage I and those with no lymph node metastasis averaged 41.7% and 57.9%, respectively. The 5-year postoperative cumulative survival rate (5Y-PCR) was less than 50% in almost all these reports. Similar data were shown in the 7 collective reviews. (2) Another 33 cases of small pancreatic cancer of 1 cm or less in diameter were collected from three reports. The rates for stage I tumor and 5Y-PCR at one institution with two reports were 100% and 100% and the rates in the other report were 85% and 78%, respectively. (3) Twelve cases of in situ carcinoma and intraductal carcinoma of the pancreas were collected from four reports. All of the patients were stage I and were alive with no evidence of tumor recurrence for periods ranging from 6 to 78 months. Small pancreatic cancer less than 1 cm in diameter is better viewed as an early pancreatic cancer, and in situ carcinoma and intraductal carcinoma of the pancreas with minimal invasion to the pancreatic parenchyma may be defined as early pancreatic cancer, regardless of size.

Abstract: We evaluated the effects of smoking on testicular function and fertilizing potential in rats. Twenty rats (group A) were exposed to the smoke of 20 cigarettes for 1 h per day. Ten rats (group B) were exposed to the smoke of 40 incense sticks for 1 h per day, and an additional 10 rats served as a control group (group C). After 10 weeks of daily exposure, serum levels of nicotine and cotinine were assessed, and a mating test was conducted. Five days later, serum concentrations of testosterone before and after human chorionic gonadotropin (hCG) stimulation, gonadotropins, and epididymal sperm content and motility were evaluated. In addition, in vitro fertilization was carried out. Nicotine and cotinine were detected in group A, but not in groups B and C. Basal serum testosterone and gonadotropin concentrations did not differ significantly among the three groups, but the testosterone response to hCG stimulation was significantly lower in group A than in groups B and C. Group A showed significant reductions in epididymal sperm content and motility, and in fertility in vivo and in vitro. These findings suggest that smoking leads to a secretory dysfunction of the Leydig cells, and also a deficiency in sperm maturation and spermatogenesis. In addition, smoking has a detrimental effect on sperm fertilizing potentials in vivo and in vitro.

We report the results of a retrospective study conducted on patients who developed recurrence following surgery for colorectal cancer, and present a practical plan to improve the survival rates. Of 974 patients who underwent surgery for colorectal cancer in our hospital during the 20 years between 1974 and 1993, 152 developed recurrence, as local recurrence in 70 (7.2%), as liver metastasis in 47 (4.8%), and as pulmonary metastasis in 35 (3.6%). The number of patients who underwent reoperation or curative resection was 54 (77.1%) and 13 (24.1%), respectively, of those with local recurrence, 16 (34.0%) and 6 (37.5%) of those with liver metastasis, and 6 (17.1%) and 6 (100.0%) of those with pulmonary metastasis. The 3- and 5-year survival rates were 13.4% and 8.9%, respectively, after reoperation for local recurrence, 14.4% and 0%, after reoperation for liver metastasis, and 53.3% and 53.3% after reoperation for pulmonary metastasis. Although evaluation was difficult owing to the small number of patients with lung metastasis, resection of a pulmonary lesion resulted in a good outcome. Furthermore, a significant difference in 3- and 5-year survival rates was observed between patients who underwent reoperation and those who underwent conservative treatment, for whom the survival rates were 0% and 0% following local recurrence, 4.6% and 0% following liver metastasis, and 0% and 0% following lung metastasis, at 3 and 5 years, respectively. These findings indicate that aggressive surgery for recurrence may result in a better outcome.

The authors recently treated a case of aggressive angiomyxoma occurring in the left scrotum. A 61-year-old Japanese man was diagnosed as having an inguinal irreducible hernia that descended to the bottom of the scrotum preoperatively. However, during the operation a large tumor was found between the spermatic fascia and the skin pressing the testis upward. The tumor measured 20×15×15 cm, was yellowish with a smooth surface encapsulated by a membrane similar to the pleura, and the cut surface was gelatinous. The histological findings indicated that the tumor had thick-walled vessels which had prominently increased in size, delicately waved collagen fibers, and stromal cells in a myxoid background. The patient has remained recurrence-free as of 11 months after surgery.

To improve the procedure of photometric determination of iron in human serum, Torii’s method recently proposed for the determination of iron was applied to serum. After the removal of protein from serum, the iron in serum is reduced, the pH of the solution is adjusted to 4.5, the color-developing reagent is added, and the excess of the reagent is removed by extraction with carbon tetrachloride. The green solution thus obtained is effective for spectrophotometric determination.The interference of copper is avoided by the addition of thiosulfate, and the substances other than copper remaining after the removal of protein from serum do not give any appreciable interference.