PACAP receptor (PAC1-R) expression in rat and rhesus monkey thymus.

The expression of PACAP receptor (PAC1-R) was investigated in the thymus of rats and rhesus monkeys. In the rat thymus, PAC1-R positive cells were found in the intermediate type of thymic epithelial cells of the medulla. PAC1-R-positive cells were also seen in the thymic medulla of the rhesus monkey. The thymus showed unusual structures in some rhesus monkey dams (F0) and offspring (F1) exposed to 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD). Additionally, in these rhesus monkeys, PAC1-R expression was different from that in the control thymus.

Immunohistopathological study of the oral lichenoid lesions of chronic GVHD.

BACKGROUND: Chronic graft-vs.-host disease (cGVHD) is a common and serious complication after bone marrow transplantation (BMT). However, the detailed process of oral lichenoid lesions of cGVHD is still unknown. Therefore, we investigated the immunohistopathological features of cGVHD compared with oral lichen planus (OLP) and healthy controls. METHODS: Nineteen allogenic BMT recipients with a histopathological diagnosis of cGVHD were investigated. We investigated the immunohistopathological features of cGVHD compared with OLP and healthy controls. RESULTS: Immunohistopathological features showed that the infiltrations of CD4-positive T cells of cGVHD and OLP were significantly larger than those of the normal oral mucosa (P < 0.005). A larger number of CD8-positive T cells was infiltrated in cGVHD and OLP compared with the normal oral mucosa (P < 0.001). The difference in the number of CD4- and CD8-positive T cells between cGVHD and OLP was not significant. The infiltrations of Langerhans cells (CD1a) in cGVHD and OLP were significantly larger than those in the normal oral mucosa (P < 0.005). The difference in the number of Langerhans cells between cGVHD and OLP was not significant. CD68-positive macrophages were more frequently seen in cGVHD and OLP than in the normal oral mucosa (P < 0.0001). The difference in the number of CD68-positive macrophages between cGVHD and OLP was not significant. CONCLUSIONS: It is suggested that Langerhans cells and CD8-positive T cell may play a major role in the pathogenesis of the oral lichenoid lesions of cGVHD, and the immune response was inducted in OLP as well as the oral lichenoid lesion of cGVHD in this study.

Hand-assisted laparoscopic nephrectomy.

PURPOSE: We report our experience in performing a hand-assisted laparoscopic radical nephrectomy (HALRN) for renal cell carcinoma (RCC) and a hand-assisted laparoscopic live donor nephrectomy (HALDN) for renal transplantation. PATIENTS AND METHODS: The clinical data on 21 patients with HALRN for stage T1N0M0 RCC and 5 living kidney donors with HALDN for renal transplantation were reviewed. We compared the results of 21 HALRNs with those of 15 conventional open radical nephrectomies for stage T1N0M0 RCC. RESULTS: The HALRN for RCC was successfully performed without any major complications in all 21 patients. The mean operation time for the HALRN group was significantly longer than that for the open surgery group (262 minutes versus 219 minutes). However, the mean estimated blood loss in the HALRN group was less than that in the open surgery group (250 ml versus 388 ml), although there was no statistically significant difference. The frequency in usage of analgesia postoperatively in the HALRN group was significantly less than that in the open surgery group (2.5 times versus 7.3 times). The HALDN was also successfully performed without any major complications in all 5 cases. The mean operation time, blood loss and warm ischemic time were 248 minutes, 322 ml and 9.8 minutes, respectively. All donors resumed oral intake of food within 48 hours after the surgery and returned to normal, non-strenuous activity by postoperative day 8. CONCLUSIONS: Our results indicate that a hand-assisted laparoscopic nephrectomy is an effective and safe surgical procedure, and it is less invasive than an open nephrectomy.

An efficient algorithm for automatically generating multivariable fuzzy systems by Fourier series method.

By exploiting the Fourier series expansion, we have developed a new constructive method of automatically generating a multivariable fuzzy inference system from any given sample set with the resulting multivariable function being constructed within any specified precision to the original sample set. The given sample sets are first decomposed into a cluster of simpler sample sets such that a single input fuzzy system is constructed readily for a sample set extracted directly from the cluster independent of the other variables. Once the relevant fuzzy rules and membership functions are constructed for each of the variables completely independent of the other variables, the resulting decomposed fuzzy rules and membership functions are integrated back into the fuzzy system appropriate for the original sample set requiring only a moderate cost of computation in the required decomposition and composition processes. After proving two basic theorems which we need to ensure the validity of the decomposition and composition processes of the system construction, we have demonstrated a constructive algorithm of a multivariable fuzzy system. Exploiting an implicit error bound analysis available at each of the construction steps, the present Fourier method is capable of implementing a more stable fuzzy system than the power series expansion method of ParNeuFuz and PolyNeuFuz, covering and implementing a wider range of more robust applications.

A new reactive nucleoside analogue for highly reactive and selective cross-linking reaction to cytidine under neutral conditions.

We have already demonstrated that the oligonucleotides DNA (ODNs) bearing a 2-amino-6-vinylpurine derivative (1) exhibited efficient interstrand cross-linking to cytidine selectively. In this study, a new reactive nucleoside analogue, 2-amino-6-(1-ethylsulfoxy)vinylpurine derivative (7), was designed based on a computational method to achieve high and selective alkylation with cytidine under neutral conditions. It has been demonstrated that the ODN (13) bearing 2-amino-6-(1-ethylsulfoxy)vinylpurine achieved highly selective and efficient cross-linking to cytidine under neutral conditions.

Hand-assisted laparoscopic live donor nephrectomy using newly produced LAP DISC: initial three cases.

BACKGROUND: A new abdominal sealing device, called the LAP DISC, was used for the first time in hand-assisted laparoscopic live donor nephrectomy (HALDN) on three donors. The LAP DISC is made of three layers of rings connected by a rubber membrane, which covers the peritoneum and abdominal wall. The upper ring can adjust to the surgeon's hand size for insertion. METHODS: The LAP DISC was seated through an approximately 7-cm midline incision under the xiphoid process. The laparoscopic port was inserted through the LAP DISC, and thereafter, pneumoperitoneum was established. Three trocars were then placed under direct vision. The surgeon's left arm was inserted into the LAP DISC and used for manual retraction, dissection, and hemostasis. In the three operations, the kidneys were removed through the LAP DISC. RESULTS: The total warm ischemic times of the kidney were 15, 8, and 4 minutes, and the total operative times were 323, 195, and 240 minutes, respectively. After the subsequent transplantation into the recipient, the kidneys produced clear urine immediately on reperfusion. The recipient creatinine fell to 4.2, 5.6, and 3.9 mg/mL on postoperative day 1. All three donors resumed consistent oral intake within 24 hours after surgery and returned to normal, nonstrenuous activity by postoperative day 6. CONCLUSION: The LAP DISC device is excellent for HALDN and may increase the number of surgeons and donors who select HALDN.

Site-directed alkylation to cytidine within duplex by the oligonucleotides containing functional nucleobases.

We have previously described that oligonucleotides (ODN) containing phenylsulfoxide derivative of 2-amino-6-vinylpurine nucleoside analog (1) are activated within duplex to form cross-link toward cytidine selectively at the target site. In this paper, we wish to report the search for more stable precursor susceptible for activation within duplex.

Efficient cross-linking to cytidine under neutral conditions using the 2-amino-6-vinylpurine derivative having an additional activating group.

We have previously reported that the oligonucleotides incorporating 2-amino-6-vinyl purine analog (1) showed efficient interstrand cross-linking with selectivity toward cytidine at a target site. In this paper, we describe the design of more efficient cross-linking reagent (2) bearing 2-amino-6-vinyl purine motif to react to cytidine selectively under neutral condition.

[Long-term results of right ventricular outflow reconstruction with valved conduit].

Twenty-nine patients who underwent right ventricular outflow tract reconstruction using a valved conduit 37 times and survived surgery, were reviewed in this study. Hancock, Ionescu-Shiley, Carpentier-Edwards and Polystan was used for RVOTR. Freedom from reoperation rates of Hancock, Ionescu-Shiley and Carpentier-Edwardsat at five and ten years after surgery were 100%, 85.7%, 89.2% and 75%, 28.6%, 89.2%, respectively. The freedom from reoperation rates of Hancock and Carpentier-Edwards were significantly higher than that of Ionescu-Shiley. In conclusion, the freedom from reoperation rate at ten years was high for both Hancock and Carpentier-Edwards.

Laparoscopic adrenalectomy for pheochromocytoma: comparison with open adrenalectomy and comparison of laparoscopic surgery for pheochromocytoma versus other adrenal tumors.

OBJECTIVE: To compare the efficacy of laparoscopic adrenalectomy for pheochromocytoma with that of conventional open adrenalectomy for pheochromocytoma and laparoscopic surgery for other adrenal tumors. PATIENTS AND METHODS: Fifty-four patients with adrenal tumors, including 10 cases of pheochromocytoma, 18 cases of Cushing's syndrome, 20 cases of primary aldosteronism, and 6 cases of nonfunctioning tumors, were evaluated. A historical group of 7 consecutive patients who underwent conventional open adrenalectomy for pheochromocytoma was also studied. RESULTS: Laparoscopic adrenalectomy for pheochromocytoma was successful in 9 of the 10 patients. There was no difference in tumor size, operation time, estimated blood loss, or occurrence of hypertensive episodes during surgery between patients treated with laparoscopic procedures and those treated with open surgery. However, the number of days to first postoperative oral feeding and first ambulation, length of hospitalization, and number of patients requiring parenteral analgesics were significantly smaller after laparoscopic surgery than after open surgery. There was no significant difference in operation time, estimated blood loss, incidence of intraoperative complications, or postoperative recovery between patients who underwent laparoscopic adrenalectomy for pheochromocytoma and those who underwent laparoscopic surgery for other adrenal lesions. CONCLUSIONS: Laparoscopic adrenalectomy does not increase the specific risks associated with surgery for pheochromocytoma. It is a minimally invasive alternative to conventional open adrenalectomy.

Hand assisted laparoscopic radical nephrectomy for renal carcinoma using a new abdominal wall sealing device.

PURPOSE: We report our initial experience with a hand assisted laparoscopic radical nephrectomy for patients with renal carcinoma, and compare our results to those of conventional open radical nephrectomy. MATERIALS AND METHODS: The clinical data on 6 consecutive patients who underwent hand assisted laparoscopic radical nephrectomy for stage T1N0M0 renal cell carcinoma were reviewed. We performed hand assisted laparoscopic surgery using the new LAP DISC* abdominal wall sealing device. We compared the results of this procedure with those of conventional open radical nephrectomy in 12 patients with stage T1N0M0 renal cell carcinoma. RESULTS: The hand assisted laparoscopic radical nephrectomy for renal carcinoma was successfully performed without any major or minor complications in all 6 patients. Mean operation time for the laparoscopic group was significantly longer than that for the open surgery group (303 minutes versus 224 minutes, p = 0.0042). However, no significant difference was observed in mean estimated blood loss for the 2 groups (264 ml. in the laparoscopic group versus 341 ml. in the open surgery group). The frequency of parenteral analgesia postoperatively in the laparoscopic group was significantly lower than that in the open surgery group (16.7% versus 75.0%, p = 0.043). In addition, the laparoscopic group seemed to recover more rapidly than the open surgery group. The abdominal wall sealing device was easy to attach to the abdominal wall, and allowed rapid hand removal and reinsertion. CONCLUSIONS: Our preliminary results indicate that a hand assisted laparoscopic radical nephrectomy with the abdominal wall sealing device is an effective and safe surgical procedure, and is less invasive than open radical nephrectomy.

Semiquantitative morphological analysis of stromal cells in the irradiated and recovering rat thymus.

To understand the roles of thymic stromal cells in T-lymphocyte development, we semiquantitatively analysed rat thymi recovering from irradiation (6 Gy), using a transmission electron microscope. The most striking findings were that the percentage of subcapsular epithelial cells significantly increased in the cortex on day 3 after irradiation compared with the control; the percentage of intermediate epithelial cells significantly increased in the cortex on days 3 and 5 after irradiation and in the medulla on days 5 and 7 compared with the control; the interdigitating cells disappeared from the medulla by day 7 after irradiation and reappeared on day 9. The present data thus reveal that during recovery after irradiation (6 Gy), marked changes occur in the relative proportions of different epithelial cell subtypes in the cortex and medulla of the rat thymus. In addition, the percentages of macrophages and interdigitating cells also changed during the recovery. These changes, which may be associated with the abrupt proliferation of thymocytes after irradiation, should shed light on the significance of stromal cells in the T cell development.

Calcitriol therapy modulates the cellular immune responses in hemodialysis patients.

We studied the in vitro and in vivo effects of calcitriol (1,25D) on the cellular immune responses in 19 hemodialysis (HD) patients. In vitro 5-day treatment with 1,25D markedly reduced the HLA-DR expression by peripheral blood CD14(+) monocytes from both HD patients and normal subjects in a similar fashion. The HLA-DR expression by monocytes and the phorbol myristate acetate (PMA)-induced superoxide production (SOP) by neutrophils were significantly higher in the HD patients than in the normal subjects (p < 0.01 and p < 0.05, respectively). The phagocytic activity in the HD patients was significantly lower than that in the normal subjects (p < 0.05). Moreover, the mitogen response of HD peripheral blood lymphocytes against pokeweed mitogen (PWM) was significantly lower than that of the controls (p < 0.01) but was only slightly and insignificantly lower against Con A. Oral 1,25D pulse therapy resulted in a marked decrease in the HLA-DR expression by peripheral blood monocytes 2 and 4 days after the first 1,25D administration (p < 0.01) in HD patients. Moreover, the treatment significantly enhanced the PMA-induced SOP 2 days after the treatment (p < 0.01). However, the phagocytic activity by neutrophils and the mitogen responses to Con A and PWM by lymphocytes were not significantly affected by this treatment. These results suggest that 1,25D plays a significant role in the regulation of both the monocyte and neutrophil functions in HD patients.

Antitumor activity of interleukin-12 against murine bladder cancer.

PURPOSE: We investigated the antitumor activity of interleukin-12 (IL-12) against MBT-2, a murine bladder carcinoma, to clarify whether or not IL-12 is effective against urothelial tumors. MATERIALS AND METHODS: MBT-2, a murine carcinogen-induced, poorly differentiated transitional cell carcinoma of C3H/He origin, was used. Three or 10 days after the subcutaneous administration of MBT-2 cells, C3H/He mice were injected intraperitoneally with IL-12 five times per wk. for 2 wk. Tumor growth was measured twice weekly. Spleen cells from the C3H/He mice that had rejected MBT-2 after the IL-12 treatment were examined for MBT-2-specific cytolytic T lymphocytes (CTL) activity and cytokine production. RESULTS: Tumor growth and acceptance was obviously suppressed when C3H/He mice were treated with IL-12 from 3 days after the tumor inoculation. In the spleen cells from the C3H/He mice that had rejected MBT-2, MBT-2-specific CTL activity and secretion of IL-2 and interferon (IFN)-gamma were clearly detected. However, the established MBT-2 tumor cells were not rejected when C3H/He mice were given IL-12 from 10 days after the tumor inoculation, although the tumor growth was transiently suppressed during the IL-12 treatment. CONCLUSION: These data demonstrate that IL-12 is considerably effective against murine bladder cancer and suggest the clinical application of IL-12 against human bladder cancer.

Interleukin-1 receptor antagonist mRNA expression and the progression of gastric carcinoma.

Interleukin-1 receptor antagonist (IL-1ra), an endogeneous inhibitor of IL-1, plays an immunosuppressive role in vivo by blocking the proinflammatory effects of IL-1. In the present study, we examined whether IL-1ra expression in human gastric carcinoma correlates with tumor progression and/or metastatic potential. The reverse transcription-polymerase chain reaction was used to compare the expression of the secreted form of IL-1ra (sIL-1ra) and the intracellular form of IL-1ra (icIL-1ra) mRNA in tumor and corresponding benign tissue obtained from 38 patients with gastric carcinoma. The incidence of sIL-1ra mRNA expression was significantly higher in tumor (52%) than in corresponding benign tissue (18%) (P = 0.002). On the contrary, icIL-1ra mRNA was detected in all tumors and benign tissues. The expression of sIL-1ra mRNA by malignant tissue correlated positively with both lymph node metastasis (P = 0.008) and liver metastasis (P = 0.015). There was no association between tumor sIL-lra mRNA expression and other clinicopathologic factors. The degree of regional lymph node reaction, such as sinus histiocytosis, in tumors expressing sI-1ra mRNA was significantly weaker than that in tumors without sIL-1ra mRNA expression (5/20 vs. 12/18, P = 0.010). These results demonstrate that the altered expression of sIL-1ra by malignant tissue may be related to the progression of gastric carcinoma via modulating host immune response.

[Chlamydia trachomatis infection in male infertility--the clinical usefulness of the detection of antibodies against Chlamydia trachomatis].

BACKGROUND: The objective of this study is to investigate the clinical usefulness of the detection of antibodies against Chlamydia trachomatis (C. trachomatis) in potential male infertility patients. PATIENTS AND METHODS: Analysis of serum samples for the presence of antibodies against C. trachomatis by enzyme immunoassay was performed in 83 male infertility patients. RESULTS: Immunoglobulin A (IgA) and G (IgG)-antibodies against C. trachomatis were found in 16 and 10 of 83 serum samples (19.3 and 12.0%) respectively. IgA and/or IgG-antibodies against C. trachomatis were found in 20 of 83 serum samples (24.1%). Serum IgA, IgG, or IgA and/or IgG positive patients showed increased semen and urine leukocytes numbers and reduced semen volume and sperm numbers, compared with negative patients, respectively, although these effects are not statistically significant. By Spearman's correlation analysis, however, significant positive correlations with serum IgA or IgG antibody index were found in semen and urine leukocytes numbers. Significant negative correlations with serum IgA antibody index was found in semen volume. In serum IgA positive patients, only 1 of 8 first-voided urine samples was positive for C. trachomatis-DNA by PCR. Moreover, the positive rates of surum IgA antibody and cervical antigen in the 8 female partners of IgA positive patients were only 25% (2 of 8 serum samples) and 12.5% (1 of 8 cervical specimens) respectively. CONCLUSION: There was a high prevalence (24.1%) of serum IgA and/or IgG-antibodies against C. trachomatis from infertility patients. The existance of antibodies against C. trachomatis in serum samples possibly influence semen quality, suggesting that C. trachomatis infection may play an important role in male infertility.

Vascular and stromal changes in irradiated and recovering rat thymus.

To analyze the mechanisms responsible for thymocyte proliferation, maturation and migration in the thymus, the rat thymus just after, and recovering from irradiation was studied morphologically. The vascular structures of the rat thymus after a radiation dose of 6 Gy were found to be destroyed on day 3, but had recovered to almost normal by day 7, suggesting that the abrupt recovery of thymus structure after irradiation was due primarily to this change in vascular structure. Furthermore, the epithelial tissues in the thymic cortex appeared to contribute to this abrupt proliferation, and possibly to the abrupt maturation of thymocytes, while medullary epithelial tissues remained sparse and appeared inactive for a relatively long period. These findings are considered important for understanding the interrelationship between thymic epithelial cells and thymocytes with respect to thymocyte proliferation, maturation and migration.

Antiproliferative effect of calcitriol on human prostatic cancer cell lines: unrelated to the expression of major histocompatibility complex antigens or intercellular adhesion molecule-1 (ICAM-1).

BACKGROUND: In addition to its role in calcium and phosphorus metabolism, calcitriol (1,25-dihydroxyvitamin D3; 1,25-D3) demonstrates multiple effects on cell proliferation/differentiation by expressing major histocompatibility complex (MHC) antigens and intercellular adhesion molecule-1 (ICAM-1). It has recently been reported that 1,25-D3 inhibits the growth of prostatic cancer (PCa) cells. In this study we examined the effect of 1,25-D3 on both the growth and expression of HLA-ABC, HLA-DR and ICAM-1 antigens in PCa cells. METHODS: Four human PCa cell lines (PC-3, PPC-1, ALVA-41 and ALVA-101) were examined. The cell numbers were enumerated, and the effects of interferon-y (IFN-gamma) and 1,25-D3 on the expression of HLA-ABC, HLA-DR and ICAM-1 were quantitated by flow cytometry. RESULTS: A dose-dependent antiproliferative effect of 1,25-D3 was found in all PCa cells lines except ALVA-41.1,25-D3 was approximately 10 times as potent as its analogue 24,25-dihydroxyvitamin D3 in inhibiting the growth of PC-3 cells. Also, the relative inhibitory ability of these compounds paralleled the strength of their binding affinities for the 1,25-D3 receptor, indicating that the antiproliferative effect may require a receptor-ligand interaction. HLA-ABC was expressed in PC-3, ALVA-41 and ALVA-101, but not in PPC-1 cells, while HLA-DR was not expressed on any of the tested cells. IFN-gammacould enhance or induce HLA-ABC but not HLA-DR expression in the tested cells. ICAM-1 was expressed in all cells and slightly upregulated by IFN-gamma. CONCLUSION: In this study 1,25-D3 had an antiproliferative effect on 3 of the 4 examined PCa cell lines.

[Bone mineral density (BMD) of the calcaneus bone in 72 dialysis patients measured by ultrasonic bone absorptiometry].

BACKGROUND: To determine what factors contribute to and change BMD in dialysis patients. METHODS: Bone parameters, namely, osteosono-assessment index (calculated by speed of sound and transmission index) were measured at calcaneus bone by ultrasonic bone absorptiometry with an Aloka Acoustic Osteoscreener (Model AOS-100). Seventy two patients were 62. 8 +/- 10.5 years of age (mean +/- SD) (range 36-81) and 70.8% were males; the patients had received dialytic therapy for 52.7 +/- 41.7 months (range 2-205). The effects of sex, age, height, weight, postmenoposal years, dialysis duration, various blood parameters, oral phosphorus binding agent (CaCO3 dosage) and oral vitamin D3 treatment (D3 dosage) on BMD were assessed statistically. RESULT: BMD in male patients were significantly higher than those in female patients (Mann-Whitney test, p < 0.01). BMD showed significant correlations with age (Spearman's correlation coefficient r = -0.384, p < 0.01), height (r = 0.479, p < 0.00001), postmenoposal years (r = -0.692, p < 0.05), dialysis duration (r = 0.250, p < 0.01), blood Ca level (r = 0.292, p < 0.05) and CaCO3 dosage, postmenoposal years and dialysis duration were the variables that significantly correlated with the BMD (multiple correlation coefficient = 0.646).

Expression of ICAM-1 in implanted primary and metastatic squamous cell carcinomas in rats.

FF6 tumor cells are derived from a spontaneous rat squamous cell carcinoma (SCC) which originally arose in the facial skin of a DA rat. In this study, FF6 tumor cells were implanted into rat oral mucosa to establish an ex vivo metastatic model. We analyzed the expression of intercellular cell adhesion molecule-1 (ICAM-1) in the implanted primary and metastatic FF6 tumors by immuno-staining with a monoclonal antibody (mAb) against ICAM-1. The implanted primary FF6 cells showed strong expression of ICAM-1, whereas the tumor cells of metastatic lesions showed weak or negative expression of ICAM-1. By immunostaining with mAb OX6, a number of MHC class II-positive macrophages were detected in tumor mesenchyme and surrounding the metastatic foci. These results suggested that the local immune reaction in the lymph node influenced the expression of ICAM-1 on tumor cells, and that MHC class II-positive macrophages may play a role in transplanted tumor growth and metastases.