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Objectives: Colorectal laterally spreading tumors (LSTs) are widely recognized owing to their structural characteristics. This study aims to clarify the histopathological characteristics of large colorectal LSTs according to growth pattern. Methods: We studied 297 colorectal LSTs measuring ≥20 mm in diameter. The LSTs were classified into four types: granular homogenous type (LST-G-H), granular nodular mixed type (LST-G-M), non-granular flat elevated type (LST-NG-F), and non-granular pseudo-depressed type (LST-NG-PD). Retrospectively collected data were examined to compare the histopathological characteristics of LSTs according to the growth pattern. Results: LST-G-M lesions (142 lesions) were most common, followed by LST-NG-F (74 lesions), LST-G-H (61 lesions), and LST-NG-PD (20 lesions). The mean tumor diameter of LST-G lesions (38.5 ± 17.2 mm) was significantly greater than that of LST-NG lesions (26.3 ± 7.0 mm, P < 0.001). In particular, 45% of LST-G-M lesions were ≥40 mm in diameter. Adenomas accounted for 54% of LST-G-H lesions compared with only 10% of LST-NG-PD lesions. Pathological T1 carcinomas accounted for 55% of LST-NG-PD lesions and were not found among LST-G-H lesions. Conclusions: The biological malignancy of colorectal LSTs differs considerably depending on the growth pattern even among large lesions and therefore should be considered when selecting treatment regimens.

Aim Portopulmonary venous anastomoses (PPVA) are shunts between esophageal varices and pulmonary veins. Because PPVA can cause serious complications at the time of sclerotherapy for esophageal varices, it is essential to confirm the existence of any PPVA before treatment. Methods The study group comprised 101 patients in whom hemodynamics were evaluated on three-dimensional computed tomography (3D-CT) before either elective or prophylactic treatment of esophageal varices at Kitasato University East Hospital from October 2007 through August 2013. The presence or absence of PPVA, laboratory test results and 3D-CT findings were retrospectively examined in these patients. Results Nine patients had PPVA, and 92 patients did not. The underlying diseases in the PPVA group were: hepatitis C liver cirrhosis in three; non-B, non-C liver cirrhosis in three; non-alcoholic steatohepatitis in one; primary biliary cirrhosis in one; and autoimmune hepatitis in one. The distribution of underlying diseases did not differ between the PPVA group and the non-PPVA group. When the study variables were statistically compared between the groups, the incidence of large, coil-shaped esophageal varices (grade F3) differed significantly between the groups (P = 0.001). Multivariate analyses of factors related to PPVA revealed that only the grade F3 type of esophageal varices differed significantly between the groups (P = 0.005; hazard ratio, 5.21; 95% confidence interval, 3.1–16.4). Conclusion In patients with grade F3 esophageal varices, the treatment method should be selected on the basis of an accurate hemodynamic analysis using 3D-CT before therapy.

Minimal deviation adenocarcinoma (MDA) is defined as an extremely well differentiated variant of endocervical adenocarcinoma. Several reports have stated that MDA associates with lobular endocervical glandular hyperplasia (LEGH). It is difficult to distinguish LEGH from MDA based on clinical and histologic similarities. There is no definite evidence proving that LEGH is a precursor lesion of MDA. A 45-year-old woman was admitted to our hospital for minute investigation of her neurological disorder. The multiple-cystic lesion at the uterine cervix was identified by magnetic resonance imaging. Based on her normal histological findings and severe underlying conditions, a careful follow-up strategy was adapted. Two years later, atypical glandular cells were observed and the multiple-cystic lesion had increased. Pathological diagnosis of a conization specimen was MDA. Radical hysterectomy was carried out. Pathological examination revealed coexistence of LEGH and MDA. Her clinical course and histological findings suggested the possibility that LEGH might be a precursor lesion of MDA.

Aim: We evaluated the long-term effects of olmesartan, an angiotensin type 1 receptor blocker, in patients with non-alcoholic steatohepatitis (NASH) complicated with hypertension. Methods: All patients were given a standard calorie diet and exercise counseling more than 3 months before the treatment. Seven patients with NASH received olmesartan treatment for 1 year. Liver biopsy, clinical parameters and blood markers of hepatic fibrosis, including serum hyaluronic acid, type IV collagen, and procollagen III N-terminal propeptide levels, were also examined at the beginning and the end of the study. Results: The median dose of the final administration was 20 mg (range, 10 - 40 mg). Olmesartan reduced MAP by –11.3 ± 13.0% (P = 0.046) after 1 year. In the laboratory data, serum AST, ALT, and ferritin significantly decreased after a year of administration (AST, 62 ± 24 vs. 39 ± 20 IU/L, P = 0.018; ALT, 106 ± 79 vs. 55 ± 35 IU/L, P = 0.043; ferritin, 323.8 ± 252.8 vs. 202.3 ± 194.1 ng/ml, P = 0.028). Furthermore, fasting glucose significantly decreased. However, transforming growth factor-beta1, the serum concentration of the fibrosis markers, and all histological features were unchanged at the end of the study. No side effects of the treatment were noted at any time during the study. Conclusion: Olmesartan significantly reduced blood pressure, fasting glucose, aminotransferase, and serum ferritin but could not suppress the hepatic fibrosis markers or histological features after 1 year. Therefore, olmesartan is advisable only for its anti-inflammatory effect in patients with NASH-complicated hypertension.

Residual symptoms are common in depression, and their presence is associated with poorer clinical outcomes of depression. We conducted a case series study of first-onset major depression to elucidate the clinical course of residual insomnia and examine the relationship between residual insomnia and recurrence of depression. Subjects were 128 patients (57 males; mean age 52.8 years) with first-onset major depression. For all patients, we continuously assessed the number and breakdown of residual symptoms listed on the 17-item Hamilton Rating Scale for Depression and quantities of prescribed psychotropic medications during the depressive and remission phases. Even during the first remission phase, 85.9% of the patients with first-onset major depression experienced an average of 2.95 residual symptoms. The most common residual symptom was insomnia (65.4%), followed by reduced work and interests (43.3%) and fatigue (39.4%). Each additional recurrence resulted in a significantly shorter remission phase as well as significant increases in antidepressant and hypnotics dosages. Hypnotics dosage during the first remission phase for patients with three or more recurrent episodes was significantly higher than that for those with only a single episode. Our findings suggest a possible link between treatment-resistant residual insomnia during the first remission phase and recurrence risk of depression. In particular, it is possible that presence of treatment-resistant insomnia during the first remission phase is related to later recurrence of depressive episodes. It is important to see patients with treatment-resistant insomnia of early stage carefully, with special attention to treatment adherence.

Aim: To determine whether concurrent chemoradiotherapy (CCRT) can improve the survival rate of high-risk uterine cervical cancer. Material & Methods: We analyzed 16 cases of uterine cervical cancer that had undergone radical hysterectomy and pelvic lymphadenectomy from 2003 to 2008. The patients were eligible if they had histologically confirmed positive parametrial involvement, positive pelvic lymph nodes or non-squamous cell carcinoma. They received 50 Gy of external beam radiotherapy (RT) for the pelvis which was combined with chemotherapy. Cisplatin was administered intravenously every 3 weeks at a dose of 70 mg/m2 during the RT. For renal function complication case, carboplatin was administered weekly. For control purposes, there were 14 cases treated in our hospital from 1995 to 2003 who had received only RT. Results: We did not find any statistically significant difference in the disease-free survival rate between the CCRT group and the RT group. However, the overall survival rate was significantly higher for patients in the CCRT group compared with the RT group in positive lymph node cases and non-squamous cell carcinoma cases. Adverse effects were more frequent in the CCRT group. Over grade 3 toxicities were manifested as leukopenia, diarrhea and anemia. There was no local recurrence in CCRT group patients. Conclusion: CCRT seems to be beneficial for improving the survival rate of either positive lymph node or non-squamous cell carcinoma cases in high-risk uterine cervical cancer patients.

This study focused on the design of a matrix for a cultured dermal substitute (CDS). Two types of cross-linked spongy matrix were designed. The weight ratio of hyaluronic acid (HA) to a cross-linking agent was adjusted to 5:1 (c-5) or 15:1 (c-15). Each HA spongy matrix was combined with collagen (Col) gel, forming 4 groups: Group I, c-5 sponge and Col gel; Group II, c-5 sponge and Col gel with human fibroblasts; Group III, c-15 sponge and Col gel; and Group IV, c-15 sponge and Col gel with human fibroblasts. The physical properties of the matrix depend on the cross-linking conditions. In animal tests using T-cell-deficient F344 rats, materials from Groups I-IV were transplanted onto a full-thickness skin defect, followed by auto-skin grafting. Commercially available artificial dermis composed of collagen sponge was employed as a control. Grafted skin survival in all groups was evaluated according to macroscopic and histological appearances 2 weeks after transplantation. The take rate of auto-skin was higher in Groups I-IV than in control. The take rate of auto-skin was also higher for Groups II and IV than for Groups I and III. This finding indicates that CDS containing human fibroblasts is able to provide more suitable conditions for rat's auto-skin engraftment. The take rate of auto-skin in Group IV was >84%, showing the best results. This finding suggests that CDS comprising highly biodegradable matrix is acceptable for the engraftment of auto-skin, creating no physical barrier to interactions between the wound bed and auto-skin.

Several spongy wound dressings composed of various hyaluronic acid (HA) materials were developed: HA aqueous solution with cross-linking agent (epoxy compound (HAWC), without cross-linking agent (HAWOC), and combined bilayer material HAWC and HAWOC (2LHA). The microstructure, sensitivity to hyaluronidase, sensitivity to free radicals, in vitro cytotoxicity, healing potential on full-thickness defect on and diabetic ulcers were evaluated. Scanning electron micrograph views of all HA wound dressings showed an interconnected channel and high porosity structure. HAWOC was degraded rapidly in hyaluronidase and free radical solution. The in vitro cytotoxicity of HAWC was significantly (p

In this study, we report a tumor that resembled previously reported uncommon tumors histologically similar to ameloblastoma or adenomatoid odontogenic tumor (AOT), showing the formation of hard tissue. We evaluated the histological characteristics by reviewing the literature. The patient was a 19-year old male. The lesion was located from the canine to third molar in the right mandible and was unicystic with a comparatively clear demarcation. The tumor tissue was cystic overall, showing multiple formation of small and large cysts. The tumor tissue resembled a variant form of plexiform ameloblastoma. Formation of dentin and dentinoid was observed in the tumor stroma, whereas formation of enamel was not observed. Very few cases of a variant form of ameloblastoma that shows formation of dentinoid have been reported, and the histological picture in this study closely resembled previously reported "adenoid ameloblastoma with dentinoid".

Background: Sarcoid uveitis is occasionally accompanied by proliferative changes, such as retinal neovascularization and vitreous hemorrhage. Steroid administration, retinal photocoagulation, and vitrectomy may be indicated in such proliferative cases. Case: A 19-year-old woman presented with proliferative sarcoid uveitis accompanied by recurrent vitreous hemorrhage. Observations: At the initial examination, bilateral vitreous opacity, retinal exudates, mild vitreous hemorrhage, retinal vasculitis, and neovascularization of the retina and optic disc were observed. Although prednisolone was administered and panretinal photocoagulation was performed several times, recurrent vitreous hemorrhage continued. Since the vitreous hemorrhage was not absorbed, pars plana vitrectomy and lensectomy were performed. After surgery, neovascularization and intraocular inflammation decreased, and the corrected visual acuity in the right eye improved to 20/50. Histopathologic analysis of the proliferative membrane removed during surgery revealed substantial neovascularization and numerous neutrophils in the vessels. Conclusions: Based on these findings, an inflammatory reaction as well as retinal ischemia were thought to be involved in the proliferative changes in this patient.

Background: There is growing evidence that the dysregulation of circadian rhythms may play an important role in irregular sleep–waking in demented elderly. In this study, we investigated daily variation of the pineal hormone melatonin, which has been reported to possess hypnogenic and synchronizing effects, in patients with senile dementia of Alzheimer’s type. Methods: Serum melatonin secretion rhythms in inpatients with senile dementia of Alzheimer’s type (SDAT group, n = 10, average age = 75.7 years) with disturbed sleep–waking and nondemented elderly (ND group, n = 10, age = 78.3 years) without clinical sleep disorders in the same facility were monitored under a dim light condition without excessive physical exercise. Results: The SDAT group showed a significantly higher degree of irregularities in actigraphically recorded rest–activity (R-A) rhythm during the 7-day baseline period compared with the ND group. The SDAT group simultaneously showed significantly reduced amplitude, larger variation of peak times, and diminished amount of total secretion in the melatonin secretion rhythm compared with the ND group. There were significantly positive correlations between the severity of R-A rhythm disorder and the reduced amplitude as well as diminished amount of total melatonin secretion. Conclusions: The SDAT patients with disturbed sleep–waking possessed melatonin secretion rhythm disorders that may play an important role in irregular sleep–waking in demented elderly.

Endothelin (ET) is well known to be a potent vasoconstrictor peptide with autocrine and paracrine function. It has been documented that ET is also present in non-muscle tissues. The distribution of ET and ET-A receptor (ET-AR) in the monkey lacrimal gland was investigated by immunohistochemistry. Three adult male monkeys (Macaca fuscata) were perfused with a fixative. The lacrimal glands were then dissected and sectioned. Using rabbit anti-ET and anti-ET-AR antibodies, the immunohistochemical procedure was performed following an ABC technique. Some sections were treated with rhodamine-phalloidin, which selectively binds to actin filaments. ET immunoreactivity was present in stellate-shaped cells located around the alveoli. In sections double-stained with anti-ET antibody and rhodamine-phalloidin, ET immunoreactivity and abundant filamentous actin were identified in the same stellate cells. Immunostaining for ET-AR was also found in the stellate shape cells. The configuration of, and the abundance of actin filaments in the stellate-shaped ET- and ET-AR immunoreactive cells suggest that they are myoepithelial cells, which are contractile and may contribute to the process of lacrimal gland secretion or maintenance of the contour of the glandular endpieces. Our results indicate that endothelin is present in myoepithelial cells of the monkey lacrimal gland.

This study investigated the cytokine expression profile in alkali-burned mouse corneas, in order to elucidate the mechanisms of corneal damage and repair.The cytokines expressed in alkali-burned corneas were identified by polymerase chain reaction (RT-PCR), then quantitated using ELISA. Based on the ELISA results, immunohistochemical analyses were performed to localize cytokine expression.Among the ten cytokines examined, IL-1 (IL-1 beta), IL-6, IL-10 and TNF-alpha mRNA were expressed in alkali-burned corneas. Quantitation revealed that IL-alpha and IL-6 were strongly induced in the early stages of alkali burn, peak production of IL-1 alpha (53.2 pg/cornea) and IL-6 (23.6 pg/cornea) occurring at days 3 and 7, respectively. The production of IL-10 and TNF-alpha was not significantly elevated during the 42 day period after burn. Immunohistochemical analyses revealed that both IL-1 alpha and IL-6 were mainly localized in regenerating epithelial basal cells.IL-1 alpha and IL-6 levels in the cornea are dramatically elevated in the regenerated epithelium during the early stages of alkali burn, and may play an important role in associated corneal damage and repair.

Purpose To investigate the mechanism of atopic cataract development, we examined a tissue-destroying major basic protein, derived from eosinophils, in the anterior capsule of cataractous lenses and the aqueous fluid of atopic dermatitis patients. Methods Major basic protein deposition in anterior capsule tissues was evaluated immunohistochemically using anti-major basic protein monoclonal antibody in three consecutive eyes from three atopic cataract patients and three consecutive eyes from three senile cataract patients. Also, major basic protein concentration in aqueous fluid obtained during cataract surgery was measured by radioimmunoassay in 12 consecutive eyes from 11 atopic cataract patients and 15 consecutive eyes from 15 senile cataract patients. Results Immunohistochemical investigation demonstrated lens epithelial major basic protein deposition in all three atopic cataracts but not in any of the senile cataracts. Major basic protein was detected in the aqueous fluid by radioimmunoassay in four of the 12 atopic cataract eyes and ranged in concentration from 11 to 70 ng/ml. No major basic protein was detected in the aqueous fluid of the 15 senile cataract eyes. Conclusion Major basic protein in anterior capsule tissues and in the aqueous fluid of eyes with atopic cataract may be associated with the pathogenesis of atopic cataract.

To evaluate the capacity of fiber outgrowth of serotonergic and dopaminergic neurons from the dorsal raphe tissue, the following three experiments were performed; (1) fetal mesencephalic raphe tissue was transplanted into the ventricle near the denervated hippocampal formation of adult rats, (2) fetal mesencephalic raphe and neonatal hippocampal tissues were transplanted into the anterior eye chamber of adult rats, and (3) fetal mesencephalic raphe tissue was explanted together with the neonatal hippocampal tissue. The extent of the fiber outgrowth was examined immunohistochemically using serotonin and tyrosine hydroxylase (TH) antisera. Three months after transplantation into the host brain, serotonin-immunoreactive (ir) fibers from raphe graft were densely distributed throughout the graft and in the host hippocampal formation, and TH-ir fibers were restricted to an area near the somata of TH-ir neurons. In particular, hyperinnervation of serotonin-ir fibers was observed in the molecular layer of the dentate gyrus. Two months after intraocular transplantation, mesencephalic raphe tissue contained a large number of serotonin- and TH-ir neurons and fibers. The distribution pattern of outgrowing serotonin-ir fibers in the hippocampal tissue was similar to that observed following intraventricular transplantation. Two weeks after explantation, the raphe tissue contained numerous serotonin-ir neurons and their fibers. These fibers extended into the hippocampal tissue in the same manner as the intraventricular and intraocular transplants. These results indicate that the intrinsic factors of hippocampal tissue influence the organization of serotonergic fibers in the hippocampal formation.

Serotonergic neurons in the fetal dorsal raphe nucleus were grafted together with fetal anterior hypothalamic tissue including the suprachiasmatic nucleus (SCN) to the anterior eye chamber of adult rats. After 6 weeks transplantation, the double grafts were immunocytochemically examined using antisera against serotonin, arginine vasopressin (AVP) and vasoactive intestinal polypeptide (VIP). The raphe grafts contained a large number of serotonin-immunoreactive neurons and fibers, but only a few AVP-immunoreactive fibers and VIP-immunoreactive neurons and fibers. On the other hand, numerous AVP- and VIP-immunoreactive neurons and fibers were found in the SCN of the anterior hypothalamic graft. Outgrowing serotonin-immunoreactive fibers from the raphe tissue were densely distributed in the anterior hypothalamic graft. In the SCN, however, only a few fibers were detected. The results demonstrate that the isolated anterior hypothalamic grafts can be innervated by the serotonergic neurons from the raphe grafts, but the innervation pattern of these fibers was quite different from the normal rat. The present results indicate that the isolated SCN has an inhibitory influence on the growth of serotonergic fibers.

Fetal rat spinal cord transplanted into the anterior chamber of the eye of an adult rat was immunohistochemically stained using antisera to substance P (SP), neuropeptide Y (NPY), methionine-enkephalin (ENK), vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP) and tyrosine hydroxylase (TH), and distributional changes of peptide- and enzyme-containing neurons 1, 2 and 4 weeks after transplantation were investigated. To examine the effect of colchicine on immunoreactivity, unilateral eyes of these adult host rats received intraocular colchicine treatment. Without colchicine treatment, numerous SP- and CGRP-immunoreactive (IR) neurons were observed in the graft 1 week after transplantation, and their immunoreactivity gradually dereeased up to 4 weeks after transplantation. NPY-, ENK-and VIP-IR neurons first appeared in the graft 2 weeks after transplantation. Four weeks after transplantation, the immunoreactivity of NPY and ENK decreased significantly, whereas VIP-IR neurons showed the same intensity as that observed at 2 weeks after transplantation. TH-IR neurons, on the other hand, were seen at every stage, but their immunoreactivity was constant all the time. After colchicine treatment, the number of SP-, NPY-, ENK- and CGRP-IR neurons appeared to increase, while that of VIP- and TH-IR neurons did not change significantly. The distribution patterns of the peptide- and enzyme-containing fibers differed from each other. In the analysis of serial sections stained with 5 peptides (SP, NPY, ENK, VIP, CGRP), fibers containing these peptides were found to be densely accumulated in specific areas of the transplanted spinal cord. The present findings demonstrated that most of the peptide- and enzyme-containing neurons systems in the transplante spinal cord showed similar distribution patterns and development to those in the normal spinal cord, but that some displayed different distribution.

In patients with burns, bone exposure accompanies serious problems which occasionally lead to amputation. We present a case of an 82-year-old woman who sustained 22% of total body surface area flame burns on her bilateral lower extremities with bone exposure. Despite fascial excision and mesh skin graft, muscles, bones, and tendons were widely exposed on her right leg. The wound was infected by methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa. To promote wound healing, we applied an allogeneic cultured dermal substitute (CDS) to the wound surface once weekly, resulting in healthy granulation except for the exposed bone area of the right anterior tibia. We then shaved the cortex of the exposed bone surface until bone marrow bleeding, and grafted mesh skin in combination with CDS. Finally, all wounds healed without osteomyelitis. The use of CDS to treat deep burns exposing bone surface may expand reconstructive options for extremities that otherwise might have been amputated.

Skin ulcers in systemic lupus erythematosus (SLE) patients are non-healing or intractable, because various factors or complications, including vasculitis and immunosuppressants, impair wound healing. In the present study, we applied cultured dermal substitutes (CDSs) to 3 cases of SLE skin ulcers because various systemic or topical therapies were ineffective. CDSs are prepared by culturing human fibroblasts on two-layered spongy matrices of hyaluronic acid and atelo-collagen, and they effectively promote the healing of severe skin defects. After using CDSs in the 3 cases, healthy granulation tissues formed within 6 weeks, and skin grafts were successfully performed. These results indicate that allogeneic CDSs provide new therapeutic alternatives as topical therapies for intractable skin ulcers in SLE.

Plasma membrane-associated adenosine triphosphatase (ATPase) samples partially purified from the tumor dissections of 15 gastric cancer patients were examined for sensitivity to the synthetic lignan, 2,3-dibenzylbutane-1,4-diol (hattalin), and ouabain in the presence of Mg2+, Na+, and K+. Hattalin was the strongest Na+, K(+)-ATPase inhibitor among the lignans previously examined. The enzyme from normal gastric tissue of the same patient was used as control. The specific activity of ATPase from cancer tissue (C-ATPase) was inhibited by more than 50% by 2.0 mM hattalin, whereas only 33.1% of the specific activity of ATPase from normal gastric mucosa (N-ATPase) was inhibited by 2.0 mM hattalin. There was statistical significance of lignan sensitivity between C- and N-ATPase (p less than 0.02). Ouabain also inhibited C-ATPase in preference to N-ATPase, though not significantly. Hattalin inhibited both C- and N-ATPase more strongly than did ouabain (p less than 0.05). Moreover, the lignan inhibited both C- and N-ATPase in the absence of Na+ and K+. From these data, it is evident that the sensitivity of plasma membrane-associated to lignan increased by gastric canceration. The target ATPase of hattalin is likely to be one other than sodium- and potassium-dependent, ouabain-sensitive ATPase.

A 52-year-old man who had lived in Brazil from 1957 through 1994 visited our department on June 17, 1997 because of a rough palate. On presentation, a 63× 29 mm ulcerative granulomatous lesion was observed in the palate. Histopathological examination revealed yeast cells with multiple budding. Paracoccidioidomycosis was diagnosed on the basis of the patients history, histological findings, and a positive reaction for Paracoccidioides brasiliensis antigen on immunodiffusion tests.Paracoccidioidomycosis is endemic in Latin America, and all cases reported in Japan were contracted abroad. This is the 14th case of paracoccidioidomycosis reported in Japan.

Pyoderma gangrenosum (PG) is an idiopathic, inflammatory, ulcerative disease of undetermined cause. Both topical and systemic treatments of corticosteroids and cyclosporine are commonly used for the ulcers of PG, but these ulcers are often intractable despite treatment. We employed allogeneic cultured dermal substitutes (CDS) in a patient with intractable ulcers due to PG. The CDS was prepared by cultured human fibroblasts on two-layered sponges of extracellular matrix such as hyaluronic acid and atelo-collagen. In the present case, re-epithelization and healthy granulation were induced by the CDS without increasing the dosage of systemic prednisolone. Then the subsequent autologous skin graft was successfully performed. This indicates that CDS is one of the useful tools for the treatment of intractable ulcers in patients with PG.

Aims Circadian clocks regulate daily rhythms of behavior and physiology such as the sleep–wake cycle and hormonal secretion. Numerous characteristics of the behavioral and physiological processes change with age. In this study, we evaluated the circadian clockwork in older people by measuring daily profiles of PERIOD ( PER ) gene expression in peripheral blood mononuclear cells (PBMCs). Main methods Blood samples were collected from 6 healthy older subjects (mean age 62 years) at 2-h intervals over a 24-h period under a semi-constant routine condition where masking effects are minimized. PBMCs were isolated from whole blood and temporal mRNA expression profiles of PER1 , PER2 , and PER3 were determined by RT-PCR. Phases of the PER rhythms, and times of sleep onset and offset were determined using data from those subjects who showed significant 24-h rhythms. The values for the parameters were compared between the older subjects and 8 young control subjects (mean age 21 years). Key findings Prominent daily rhythms of PER1 , PER2 , and PER3 mRNA levels, advanced sleep–wake timing and advanced phases of PER rhythms were observed in the older subjects compared to the young controls. There was no significant age-related phase difference in PER1 or PER2 rhythm with respect to sleep timing; however, PER3 expression pattern was altered in the older subjects. Significance This preliminary study shows that human circadian clockwork in PBMCs remains intact at least until the presenile stage and suggests that the altered PER3 expression pattern may reflect decreased homeostatic sleep drive in older people.

The effect of each of twelve mammalian lignan derivatives on the growth of human mammary tumor ZR-75-1 cells was examined. At a concentration less than 10 micrograms/ml, tumor cell growth was inhibited from 18-68%. The effect of 2,3-dibenzylbutane-1,4-diol(hattalin) was found to be strongest, inhibiting growth by 50% at a concentration (EC50) of 2.1 micrograms/ml. Hattalin inhibited membrane Na+, K(+)-ATPase of canine kidney cortex. It also inhibited the ATPase of the plasma membrane fraction from both cultured cells and a section of human breast cancer tissue at a concentration ranging from 0.5 to 2.0 mM. However, only a few percent of membrane ATPase from either ZR-75-1 cells or breast carcinoma tissue was inhibited by 2.0 mM of ouabain, suggesting that the target ATPase of hattalin was other than ouabain-sensitive ATPase. The relative incorporation of [3H]thymidine per 1 x 10(5) cells into the acid-precipitable fraction of ZR-75-1 cells was not affected by 1-50 micrograms/ml of hattalin, while a marked decrease resulted from 1-10 micrograms/ml of 5-fluorouracil (5-FU). These results suggest that the suppressive effect of hattalin on tumor cell growth may not occur through inhibition of DNA synthesis but rather partly by inhibition of the plasma membrane ATPase other than Na+ and K(+)-dependent ones.

We report on a 7-year-old boy in whom a wooden foreign body was found in the temporal region. A wooden foreign body was not suspected initially, because there was no past history of trauma according to the patient and his parents. A thumb-sized mass, suspected to be a malignant tumor, was found in the left temporal region of the head. Plain radiography, computed tomography, and magnetic resonance imaging revealed no evidence of a wooden foreign body. Diagnosis was difficult, and the wooden foreign body was detected on a second surgical exploration.

We report on a fetus with a congenital pulmonary myofibroblastic tumor, the prenatal detection of which with imaging modalities has not been reported up until now. A 32-year-old woman was referred to our hospital at 29 weeks' gestation because of severe fetal hydrops. Sonograms and magnetic resonance imaging showed a large solid tumor in the left thorax. The fetus died in utero the next day. Autopsy confirmed that the tumor was confined to the lower lobe of the left lung, and circulatory insufficiency from compression by the tumor was considered to be the cause of fetal hydrops and demise. Histologic examination revealed that the tumor was composed of uniform short spindle cells with no atypia and a large number of vessels. In addition, with immunohistochemical studies, the tumor cells were stained for calponin but not for cluster differentiation (CD)-31, CD-34, alpha-smooth muscle actin or S-100.

We report two cases of fetofetal transfusion in monochorionic triamniotic triplet pregnancies. Case 1: At 23 weeks' gestation an amnioreduction was carried out. Three days later, the donor triplet died in utero. Immediately after that, a cardiotocogram of the triplet who had been thought to be unaffected by the fetofetal transfusion, showed a non-reassuring fetal status. Although cesarean section was carried out, none of the triplets survived. Case 2: At 24 weeks' gestation a woman was transferred to our center because of fetofetal transfusion in monochorionic triplets. Cesarean section was carried out. However, the recipient died on the 75th day after birth, and the others had neurological problems. Previous reports on fetofetal transfusion in triplets are very limited. The prognosis of this condition has been reported to be severe, irrespective of chorionicity, gestational age at delivery, maternal age, and parity. When managing a monochorionic or dichorionic triplet pregnancy, serial and careful ultrasound examination is mandatory to find early symptoms of this serious condition.

We report two cases of a fetus with sirenomelia sequence which showed oligohydramnios and a single umbilical artery. The first case was of a single fetus with symelia apus and only one leg. Prenatal diagnosis of this case was possible. The second case was of a dichorionic-diamniotic twin pregnancy in which one fetus had symelia dipus with two fused lower extremities. Prenatal diagnosis of the condition was not made. In both cases, the fetuses died shortly after birth from respiratory distress due to severe pulmonary hypoplasia. Absence of urinary tract, imperforate anus, and spine deformity were confirmed in both cases. Although prenatal diagnosis of symelia dipus seems difficult, this condition must be considered in a fetus with severe oligohydramnios.

We describe a 28-year-old man with a papillary cyst adenoma arising in the hard palate. The patient came to the hospital for further evaluation of a mass in the hard palate. MRI examination revealed a mass in the hard palate and signs of bone resorption. The mass was enucleated in a single mass under general anesthesia. The histopathological diagnosis was a papillary cyst adenoma. Forty-two months have passed since the operation, and the patient's progress has been good, without any signs of recurrence.

Osteoblastoma arises primarily in the vertebral column, limbs, and other long tubular bones. It rarely occurs in the jaws.This report describes a 22-year-old woman who visited our department because of swelling and pain at 3 2. Intraoral examination revealed marked pain on percussion at 3 2 with turgescence on the lingual side. Radiographic examination showed sequestrum-like findings with the presence of both radiopaque and radiolucent areas.Under local anestesia, 3 2 were extracted and the tumor was extirpated en bloc. Pathological examination suggested that the lesion was an osteoblastoma. The patient's postoperative course is favorable as of 41 months after the operation.

It is difficult to identify foreign bodies by simple X-ray, CT, and MRI examination if the foreign body is radiolucent or resides in a deep region.In this paper, we describe a patient with a foreign body (an ear of rice) in the left submandibular gland. Differentiation of the foreign body from simple inflammatory or tumorous lesions was difficult owing to the rare clinical characteristics of this case.

We report a case of candidiasis occurring in the maxillary sinus.The patient was a 53-year-old woman who presented with a major complaint of a strange feeling of the right cheek. Computed tomographic examination revealed an opaque, softtissue-like mass in the right maxillary sinus, which contained a high-density region. The clinical diagnosis was right chronic maxillary sinusitis accompanied by a foreign body in the maxillary sinus.Caldwell-Luc operation was performed under general anesthesia. Caseous material was found in the sinus, and it was removed. The postoperative histopathological diagnosis was candidiasis. Three years have passed since operation, and the patient's progress has been good.

Hyperplasia of the minor salivary glands is an idiopathic rare lesion;most lesions arise on the palate and appear as an asymptomatic localized tumor, which is difficult to differentiate from other tumorous lesions.Histpathologically this lesion has normal salivary glands, with no signs of inflammation.In this paper, a case of hyperplasia of the minor salivary gland (palatine gland), located on the soft palate in a 46-year-old man, is reported.

Odontogenic fibroma derived from the mesenchymal tissue of teeth is a rare benign odontogenic tumor. This tumor usually contains inactive odontogenic epithelium and occasionally involves bone-like or cementum-like tissue.We report a 38-year-old man in whom a central odontogenic fibroma developed in the region of impacted 45. The differential histopathological diagnosis of this case was most likely a cementifying fibroma.

Study objectives To investigate the effects of one night's total sleep deprivation (TSD) on NK cell activity, with rigorous control of circadian phase of sampling points as well as physical exercise level in association with sleep deprivation. Design The mean sleep onset time of each subject before starting the study was defined as his 0000 h. This study was composed of a Sleep-Sleep session (sleep times, 00:00 h - 08:00 h and 24:00 h - 32:00 h) and a Sleep-Wake session (sleep time, 00:00 h - 08:00 h) with TSD (24:00 h - 32:00 h) placed in a cross-over design with 2-week interval between each session. In each session, the subjects were rested in the supine position under dim light from - 06:00 h to 36:00 h (for 42 hours). Setting University-based sleep and chronobiology laboratory Participants 10 healthy adult men (mean age, 20.9 y; age range, 19-23 y) Interventions NA. Measurements and results NK cell activity was measured every 4 hours from 12:00 h. NK cell activity during TSD (at 28:00 h) has been revealed to significantly increase (p=0.01) compared with the corresponding value in the Sleep-Sleep session. This effect was weaker at their usual waking time 32:00 h (p=0.07), and disappeared until 36:00 h (4 hours after awakening). The circadian rhythm phases (dim light melatonin onset time) were coincident between the 2 sessions. Conclusions The present findings suggest that one night TSD induces an acute and transient increase in NK cell activity that is not influenced by the effects of circadian rhythm or the amount of physical exercise undertaken during TSD.

To investigate the long-term effect of a topically applied beta-blocker on human corneal epithelium, the corneal epithelial barrier function and the superficial cell area of the corneal epithelium were evaluated. Seventeen normal healthy volunteers (without medication), 7 cataract patients (treated with pyrenoxine eyedrops) and 7 glaucoma or ocular hypertension patients (treated with 0.5% timolol maleate) were assigned to this study. The eyedrops had been used on a daily basis for at least 3 months. In the evaluation of corneal epithelial barrier function, fluorescein uptake was measured using a slitlamp fluorophotometer after application of 3 microl of 0.5% fluorescein for 10 min. In the evaluation of the superficial cell area, the central corneal epithelium was measured by tandem scanning confocal microscopy (TSCM). The healthy control and timolol groups were compared. Corneal fluorescein uptake in the healthy control, pyrenoxine and timolol groups was 20.3 +/- 3.2, 21.5 +/- 4.0 and 76.2 +/- 30.0 ng/ml (mean +/- standard error), respectively. There was a significantly higher fluorescein uptake in the timolol group compared to the pyrenoxine group (p = 0.0088) and the healthy control group (p = 0.0055). TSCM showed no significant difference in the superficial cell areas of the corneal epithelium between the healthy control and timolol groups. beta-Blocker eyedrops decreased the corneal epithelial barrier function. Their application was not accompanied by any biomicroscopic change in the superficial cell area.

Purpose: To determine the site of rebleeding into the vitreous after vitrectomy in patients with diabetic retinopathy. Methods: We studied in detail 4 eyes of 4 patients in whom rebleeding into the vitreous followed successful vitrectomy for proliferative diabetic retinopathy. In addition, the fibrous membrane removed at surgery was studied by light and electron microscopy. Results: In these 4 eyes, the second operation revealed that the source of the vitreous rebleeding was from a fibrovascular proliferation around the scleral wounds of the initial surgery, and no other neovascularization and/or reproliferation were observed in the whole retina. Rebleeding in these 4 eyes developed at an average of 9 weeks after initial surgery. The proliferative membrane was oval in shape and expanded from the residual vitreous that had been incarcerated in the scleral wound. The proliferative membrane removed during vitrectomy was poor in cellular components and contained extracellular matrix. Blood vessels of various sizes were also present. Electron microscopy showed the membrane was rich in extracellular components and contained high and low electron density cells. These cells often had microvilli and seemed to be of epithelial origin. Conclusions: These findings show that vitreous rebleeding may develop from fibrovascular proliferation from the scleral wound created during initial surgery. The proliferated membrane showed histological similarities with the fibrovascular proliferation usually seen in the diabetic retina and may represent a type of anterior proliferation secondary to retinal ischemia.

Background: We encountered a patient with central retinal vein occlusion (CRVO) forming an epiretinal membrane (ERM) in a wide area. Generally formation of ERM is rare in CRVO. Patient and Methods: The patient was a 72-year-old female with ischemic CRVO in the right eye, who had undergone insufficient pan-retinal photocoagulation. Since ERM was formed in a wide area 7 months after the occurrence of CRVO, the patient underwent vitrectomy. Results: Although the patient was elderly, the posterior vitreum had not been detached, and ERM was observed in a wide area from the disk of the optic nerve over the vascular arcade. Since the ERM strongly adhered to the retina, incision of the membrane with vitreous scissors was required. The membrane tissue collected during the operation showed few vascular and cellular components, and consisted of ERM extracellular matrices such as collagen. Conclusions: In this patient, however, it was probable that not only barrier dysfunction of retinal vessel endothelial cells caused by elevated pressure in the retinal vein and rapid retinal ischemia but also the anatomical feature of the undetached posterior vitreum was involved in the formation of the ERM.

Trabecular meshwork cells are actively phagocytic and may function to keep the drainage pathways free of cellular debris, pigment and other material. A decrease in phagocytic capacity has been proposed in the pathogenesis of glaucoma. This study was performed to compare the phagocytic capability of the human trabecular meshwork in glaucomatous and normal human eyes. The anterior segments of 6 donors with glaucoma (primary open-angle glaucoma, POAG: 5 donors; pseudoexfoliative glaucoma, PEX: 1 donor) and 6 normal donors were placed in perfusion organ culture. During the final 24 h of culture, latex microspheres, labeled with FITC and coated with antibodies, were perfused into the anterior segments. Eyes were then fixed, the trabecular meshworks were treated with a rhodamine-labeled secondary antibody, sectioned and the number of ingested beads determined with the laser scanning confocal microscope. Nuclei were quantitated and used to calculate the phagocytic index of each eye (number of ingested beads/number of nuclei). Anterior segments of glaucomatous donors were cultured for 1-3 days, as a preliminary culture study had revealed that culture of glaucomatous anterior segments is successful in only 50% when cultured for 21 days. Specimens of normal donors were cultured for 21 days. Ingested beads appeared green and could be differentiated from noningested beads, which appeared red, using appropriate wavelengths of the laser. Bead ingestion was confirmed with electron microscopy and the use of a secondary antibody labeled with horseradish peroxidase. Ingestion rates appeared similar among all three groups of eyes: POAG, 3.8 beads/cell; PEX, 3.3 beads/cell; normal, 3.5 beads/ cell. No evidence of significant migration or loss of trabecular cells was noted. Cell counts were not significantly different: POAG, 127 +/- 40 cells/section; normals, 136 +/- 49 cells/section. In conclusion, the phagocytic ability of the trabecular meshwork appears similar between eyes with POAG and normal eyes in perfusion organ culture. Cell loss after phagocytosis was not observed in these single-exposure experiments.

• Background: Collagen has effects on cell morphology, differentiation characteristics and function. Using collagen gel culture, several studies about cell differentiation were reported. In this study, the differentiation of rabbit conjunctival epithelial cells in a collagen gel-embedding culture system was investigated by electron microscope and lectin labeling. • Methods: Rabbit bulbar conjunctival epithelial cells were cultured in type I collagen gel. After I and 2 weeks of culture, some of these cells were stained with PAS and seven kinds of lectins, and others were examined by transmission electron microscopy. • Results: The conjunctival epithelial cells cultured within collagen gel formed stratified cell layers and globules with cavities. The inner layer cells facing the cavities showed PAS and lectin staining patterns similar to those of conjunctival goblet cells in vivo, whereas the staining patterns of the outer layer cells on the collagen matrices resembled the patterns of non-goblet epithelial cells. Microvilli on the surface of the innermost cells, basement membranes beneath the outermost cells, tight junctions, adherent junctions, interdigitating folds and desmosomes between cells were identified on electron microscopic examination. • Conclusion: These results indicate that cell junction structures of the conjunctival epithelial cells are well developed in collagen gel-embedding culture systems, and that the inner layer cells have carbohydrates similar to those of conjunctival goblet cells. Culture of conjunctival epithelial cells within collagen gel is a useful model for examining differentiation of these cells.

In our previous paper, the intraocular double grafts of fetal mesencephalic raphe and suprachiasmatic nucleus (SCN) demonstrated that the serotonergic fibers from raphe tissue did not show a dense innervation of SCN [28]. To examine the influence of NPY innervation from lateral geniculate nucleus (LGN) on the development of serotonergic fibers in the SCN, fetal mesencephalic raphe, SCN and LGN tissues were transplanted together into the eye chamber of adult rat. 6 weeks after transplantation, triple grafts were immunohistochemically examined. The SCN cell cluster was recognized by vasoactive intestinal polypeptide (VIP)- and arginine vasopressin (AVP)-immunoreactive neurons and fibers. The SCN cell cluster also contained a large number of serotonin-immunoreactive fibers from raphe tissue and a moderate number of neuropeptide Y (NPY)-immunoreactive fibers from LGN tissue. The present results provide information on possible NPY-serotonin interactions in the developing SCN.

Effects of Y-20811, a thromboxane A2 synthetase inhibitor, on the arachidonic acid (AA)-induced responses were investigated in the isolated canine coronary artery superfused with blood from a donor dog and Krebs-Henseleit solution. When the coronary artery was superfused with Krebs-Henseleit solution, AA did not have any remarkable effect on its tone. In the coronary artery superfused with blood, in contrast, AA (10-100 micrograms) caused phasic constriction followed by relaxation. After denudation of the endothelium, the contractile response was augmented, and the relaxant effect was abolished. Y-20811 (1 mg/kg, i.v.), administered into the donor dog, inhibited the contraction and augmented the relaxation. Indomethacin (5 mg/kg, i.v.) and aspirin (30 mg/kg, i.v.) also inhibited the AA-induced contraction. However, indomethacin inhibited the relaxation induced by AA (10-100 micrograms), whereas aspirin inhibited the relaxation induced by a low dose of AA (10-30 micrograms). These results suggest that Y-20811 inhibits the TXA2-induced contraction and augments the production of the relaxant metabolite of AA in the coronary artery.

The effects of Y-20811, a selective inhibitor of thromboxane A2 (TXA2) synthetase, on blood flow and local levels of immunoreactive thromboxane B2 (i-TXB2) and 6-keto prostaglandin F1 alpha (i-6-keto PGF1 alpha) in the coronary artery were investigated in the canine model of coronary thrombosis. Thrombosis was induced by applying an electric current to the intraluminal surface of the coronary artery. The plasma levels of i-TXB2 and i-6-keto PGF1 alpha were measured distal to the electrode. In the control group, coronary blood flow decreased and finally stopped 207 +/- 53 min (mean +/- S.E.M., n = 5) after the start of current application. The level of i-TXB2 rose before the coronary occlusion. Coronary blood flow did not change significantly in the Y-20811-treated group (1 mg/kg i.v.). The level of i-TXB2 decreased and remained significantly lower than that in the control group. The level of i-6-keto PGF1 alpha tended to increase slightly in the Y-20811-treated, but not in the control group. The weight of the thrombus in the Y-20811-treated group was significantly less than that in the control group (P less than 0.01). These results suggest that Y-20811 prevents coronary thrombosis by the inhibition of TXA2 production around the electrically injured lumen of the coronary artery.

PURPOSE We investigated the adverse effects of beta-blocker eye drops on tears and ocular surface epithelium. METHODS We studied twenty-three eyes of twenty three glaucoma patients [10 males, 13 females: 53.8 +/- 12.2 (yrs; mean +/- standard deviation)] treated with beta-blocker eye drops for more than three months and thirty two control subjects (16 males, 16 females: 50.4 +/- 10.9). The parameters described below were compared between the groups: 1. the radius of tear meniscus curvature, 2. grades for tear lipid layer interference patterns, 3. non-invasive breakup time (N-BUT), 4. cotton thread value, 5. scores of fluorescein staining, 6. fluorescein breakup time (F-BUT), 7. scores of rose bengal staining, 8. and Schirmer I value. RESULTS The glaucoma group showed a significant decrease in the radius of tear meniscus curvature (p = 0.0007), a significantly lower distribution in the grades for tear lipid layer interference patterns (p = 0.0270), a significant difference in the scores of fluorescein staining (p < 0.0001), a significant shortening in F-BUT (p = 0.0050), a significantly higher distribution in the scores of rose Bengal staining (p = 0.0010), and a significantly smaller value in Schirmer I value (p = 0.0042). However, there was no significant difference in N-BUT and cotton thread value. CONCLUSIONS These results clearly demonstrate that the ocular surface in glaucoma patients treated with beta-blocker eye drops show dry-eye-like changes in terms of tears and ocular surface epithelium.