It has been shown that pretransplant intrathymic injection of donor antigen prolongs rat cardiac allograft survival. The purpose of the present study was to evaluate whether co-administration of FK506 could enhance the beneficial effect of intrathymic preimmunization; we also wished to clarify whether this combined treatment could be effective for renal allografts. Seven days prior to transplantation 5 x 10(7) donor splenocytes were injected into the thymus of LEW (RT1l) recipients of WKA (RT1u) cardiac or renal allografts. Using a miniosmotic pump, FK506 was administered for 7 days before transplantation at a dose of 3 mg/kg/day. Whereas intrathymic injection of donor splenocytes or subcutaneous FK506 alone showed limited (15.2 days) or no (7.3 days) prolongation of cardiac allograft survival, the combined treatment with these two modalities synergistically prolonged the survival of the cardiac allograft (> 82.0 days). This combined treatment, however, failed to prolong renal allograft survival (8.4 days). Although donor-specific prolongation of a second cardiac allograft was obtained in LEW rats with a well beating WKA heart, two of the three recipients ultimately rejected both first and second WKA cardiac allografts. Additionally, LEW rats with a long-surviving WKA heart rejected a WKA renal allograft in a normal fashion. These data demonstrate a synergistic effect of intrathymic injection of donor splenocytes and subcutaneous FK506 on cardiac, but not renal, allograft survival and suggest that quantitative or qualitative differences of immunogenicity of kidney may contribute to the rejection of renal allografts.

Animals↗

PubMedDate not supplied

Viability of gynecological malignant cultured cells after 4 degrees C storage in citrate-phosphate-dextrose solution.

For the surgery of gynecological malignant tumors, a predeposit type of autologous blood transfusion has been widely used. However, using molecular biologic techniques, malignant cells have been found in the peripheral blood of cancer patients. Therefore, to evaluate the utility of presurgical blood deposits, we studied the survival of gynecological malignant cultured cells after 4 degrees C storage in citrate-phosphate-dextrose solution. Ten cultured cell lines derived from gynecological malignant tumors were used. Mixtures of 1 x 105 cells, culture medium, and citrate-phosphate-dextrose (CPD) solution were stored at 4 degrees C. After 0, 1, 4, 8, 15, or 22 days of storage, cells were placed in 96-well microtiter plates in culture medium with 1 x 104 cells/100 micro l/well. After 24 h culture in the incubator, the survival rate was calculated from the optical density by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay. More than 10% of surviving cells were seen in nine cell lines after 4 days of storage, in seven cell lines after 8 days, in three cell lines after 15 days, and in two cell lines after 22 days. Cancer cells in presurgical blood deposits may survive a 3-week storage period at 4 degrees C in CPD solution.

Blood Preservation↗

PubMedDate not supplied

Measurement of activities in two different angiotensin II generating systems, chymase and angiotensin-converting enzyme, in the vitreous fluid of vitreoretinal diseases: a possible involvement of chymase in the pathogenesis of macular hole patients.

PURPOSE: To investigate possible involvement of chymase and angiotensin-converting enzyme (ACE) in the pathogenesis of vitreoretinal diseases, both of which are related to the production of angiotensin II. METHODS: We measured chymase and ACE activities in the vitreous in the 54 affected eyes of 54 patients who had undergone vitreous surgery for idiopathic macular holes (MH, n = 14), proliferative diabetic retinopathy (PDR, n = 14), idiopathic epiretinal membranes (ERM, n = 13), and rhegmatogenous retinal detachment (RRD, n = 13). RESULTS: Chymase activities in the vitreous from patients with MH, PDR, ERM, and RRD were 1.87 +/- 0.53, 0.06 +/- 0.04, 0.40 +/- 0.12, and 0.08 +/- 0.03 (mean +/- SE) mU/mg protein, respectively, and ACE activities in the vitreous humor were 0.18 +/- 0.09, 0.30 +/- 0.07, 0.01 +/- 0.01, and 0.03 +/- 0.02 (mean +/- SE) mU/mg protein, respectively. Chymase activity was significantly elevated in MH among these diseases (p < 0.01, Scheffe), and ACE was significantly activated in PDR compared to ERM and RRD (p < 0.05, Scheffe). CONCLUSIONS: Our results suggest that two different angiotensin II generating systems are activated in human vitreous humor; an increased activity of chymase may play a possible role in the formation of macular holes.

Aged↗

PubMedDate not supplied

Psychomotor performance during initial stage of exposure to halothane, enflurane, isoflurane and sevoflurane in mice.

1. The dose-response relationship during the pre-anaesthetic and pre-equilibrium state of four inhalation anaesthetics (halothane, enflurane, isoflurane and sevoflurane) on fixed-ratio schedule-controlled behaviour was studied in mice. 2. Animals exposed to halothane, enflurane and isoflurane showed a biphasic pattern (i.e. a significant incremental increase in psychomotor responses at low inspired concentrations (0.1-0.4%) and decrements at greater concentrations) in a dose-dependent manner. Mean peak responses (per cent of control) were 132.3 (P < 0.01 vs control), 124.0 (P < 0.05) and 116.7% (P < 0.05) at 0.1% halothane, 0.3% enflurane and 0.2% isoflurane, respectively. 3. Sevoflurane did not increase schedule-controlled behavioural responses at any concentration. 4. The effect of subanaesthetic concentrations of inhalation anaesthetics on psychomotor performance can be evaluated as changes in the response rate in schedule-controlled behaviour.

Anesthetics, Inhalation↗

PubMedDate not supplied

HER-2 codon 655 polymorphism in cervical carcinogenesis.

HER-2 codon 655 polymorphism together with human papillomavirus (HPV) types were examined in a total of 279 cervical smear samples. Forty-nine patients with high-grade squamous intraepithelial lesion had higher frequency of high-risk HPV than 167 patients with low-grade squamous intraepithelial lesion and 63 controls. There was no statistical difference in the frequencies of HER-2 Ile/Ile, Ile/Val, and Val/Val genotypes between squamous intraepithelial lesions (SILs) and controls. When the Ile/Ile genotype was compared to the Ile/Val + Val/Val genotypes, there was also no statistical difference in the genotype prevalence between SILs and controls either in 91 or 188 patients with or without high-risk HPV, respectively. These results suggest that the HER-2 polymorphism at codon 655 in cervical cell samples is unlikely to be associated with HPV status and the onset of cervical cancer in a Japanese population.

Base Sequence↗

PubMedDate not supplied

Fas gene promoter -670 polymorphism in gynecological cancer.

Single-nucleotide polymorphism at -670 of Fas gene promoter (A/G) was examined in a total of 354 blood samples from normal healthy women and gynecological cancer patients. They consisted of 95 normal, 83 cervical, 108 endometrial, and 68 ovarian cancer cases. Eighty-three patients with cervical cancer had statistically higher frequency of GG genotype and G allele than 95 controls (P= 0.0353 and 0.0278, respectively). There was no significant difference in the genotype or allele prevalence between control subjects and endometrial or ovarian cancer patients. The Fas -670 GG genotype was associated with an increased risk for the development of cervical cancer (OR = 2.56, 95% CI = 1.08-6.10) compared with the AA genotype. The G allele also increased the risk of cervical cancer (OR = 1.60, 95% CI = 1.05-2.43) compared with the A allele. Germ-line polymorphism of Fas gene promoter -670 may be associated with the risk of cervical cancer in a Japanese population.

Adult↗

PubMedDate not supplied

Effects of EGF and TGF-alpha on invasion and proteinase expression of uterine cervical adenocarcinoma OMC-4 cells.

Uterine cervical adenocarcinoma typically is an aggressive neoplasm with a propensity for early invasion and dissemination; however, the regulatory mechanism of invasive activity of cervical adenocarcinoma cells has not been fully understood. In this study, biological effects of epidermal growth factor (EGF) and transforming growth factor (TGF)-alpha on invasion and proteinase expression of human cervical adenocarcinoma OMC-4 cells were investigated. Tumor cell migration along a gradient of substratum-bound fibronectin and invasion into the reconstituted basement membrane were stimulated by 0.1-10 nM EGF and TGF-alpha in a concentration-dependent manner. Their effects on tumor cell migration were also confirmed by wound assay. The zymography of tumor-conditioned medium showed that the treatment of OMC-4 cells with EGF and TGF-alpha resulted in the increase of matrix metalloproteinase (MMP)-2 and urokinase-type plasminogen activator (uPA). Matrilysin (MMP-7), also secreted by OMC-4 cells, was not affected by these growth factors. These results suggest that EGF and TGF-alpha act as positive regulators on the invasion of cervical adenocarcinoma cells, which may be associated with their stimulatory effects on tumor cell motility and the induction of type IV collagenase and uPA secreted by tumor cells.

Adenocarcinoma↗

PubMedDate not supplied

Donor-specific unresponsiveness induced by intraportal grafting and FK506 in rat islet allografts: importance of low temperature culture and transplant site on induction and maintenance.

Previously we demonstrated prolongation of islet allograft survival in rat by administration of FK506 to the recipients. The purpose of the present study was to determine whether specific immune unresponsiveness had been induced and to determine the effects of low temperature culture of donor islets as well as the transplant site on the induction of immune unresponsiveness. At 90 days after transplantation, normoglycemic recipients bearing functional intrahepatic grafts were made diabetic again with streptozotocin (STZ) and donor specific or third party islets were transplanted either into the liver or beneath the kidney capsule. When fresh islets were used as donors in initial transplantation in conjunction with FK506, intrahepatic re-transplants of fresh islets from the donor-specific strain in the absence of FK506 maintained normoglycemia for more than 60 days, while third party transplants (n = 3) were rejected within 1 wk. In contrast to intrahepatic regrafts, all the renal subcapsular regrafts from the donor-specific strain (n = 3) were rejected with mean survival time of 12.7 +/- 6.4 days. When cultured (24 degrees C, 7 days) islets were used for initial transplantation in conjunction with FK506, re-transplants of fresh or cultured islets from the donor specific strain beneath the kidney capsule maintained normoglycemic in 3 out of 6 or all (n = 4) of the recipients, respectively. Cultured third party regrafts beneath the kidney capsule (n = 2) were rejected at 9 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMedDate not supplied

Cytologic study of the tissue repair cells of the uterine cervix. With special reference to their origin.

In order to characterize tissue repair cells (TR) of the uterine cervix and clarify their origin, exfoliated cells obtained after laser conization for early cervical lesions were examined. One specimen was first examined by Papanicolaou staining and then examined for epithelial membrane antigen (EMA) and vimentin by immunocytochemical staining, using a restaining method. The other specimen was observed by phosphotungstic acid-hematoxylin (PTAH) staining. Morphologic findings on TR were investigated together with the histologic findings on the wound healing process. TR were classified morphologically into three groups: stromal (STR), epithelial (ETR) and of unknown origin (UTR). Validity of this classification was confirmed by the findings of immunocytochemical staining with EMA and vimentin. These cells appeared one to eight weeks after laser conization. TR with relatively large nuclei, or atypical TR (ATR), appeared when each type of TR was most plentiful, at two to five weeks. Regarding the origins of each TR, cytologic and histologic findings could be considered to offer evidence that ETR originated with hyperplasia of immature cells of the squamous epithelium or reserve cells below the columnar epithelium. The presence of myofibrils in cytoplasm, demonstrated by PTAH staining for STR and some UTR, strongly suggested the possibility that these cells were myofibroblasts in granulation tissue.

Carcinoma in Situ↗

PubMedDate not supplied

Visual findings and histologic diagnosis of pelvic endometriosis under laparoscopy and laparotomy.

OBJECTIVE: To assess the diagnostic value of visual findings in the diagnosis of endometriosis from the histological point of view. STUDY DESIGN: 212 specimens from 107 patients with benign (74.8%) or malignant (25.2%) disease were obtained by biopsy or resection under laparoscopy (65 patients) or laparotomy (42 patients). Ages ranged from 19 to 62 (mean age 36.4). Visual findings were classified according to the criteria established by the Endometriosis Committee of the Japan Society of Obstetrics and Gynecology in 1993. Specimens were stained with hematoxylin-eosin, and 15 cases with periodic acid-Schiff stain or silver impregnation stain. RESULTS: Among pigmented lesions, endometriosis was found in 73.0% of specimens from the pelvic peritoneum and in 56.4% of those from the ovaries. Blueberry spots in the pelvic peritoneum as well as ovarian chocolate cysts showed the highest positive rate. In the presence of multiple or complex pigmented lesions of the pelvic area, the rate was still higher (88.6%). Those rates were due to our inclusion of inaccurate and incomplete biopsy specimens. Endometriosis of nonpigmented lesions was found in only 11 patients (12.0%) who also had pigmented lesions and/or adenomyosis. CONCLUSION: The laparoscopic diagnosis of endometriosis can be made only when multiple complex pigmented lesions are observed, but, otherwise, histopathological confirmation is necessary.

Adult↗