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Although population characteristics and antimicrobial prescribing practices suggest that the hospitalized population in Japan is at high risk of Clostridioides difficile infection (CDI), the epidemiology of CDI in Japan is poorly understood.This prospective cohort study aimed to investigate the epidemiology of CDI at 12 hospitals in Japan.Patients with clinically significant diarrhoea (CSD) were enrolled. Stool specimens were tested for C. difficile by toxin A and/or B enzyme immunoassay (EIA) in the hospital laboratories, and a toxigenic culture and nucleic acid amplification tests were performed at a central laboratory. The risk factors of CDI and the impact of CDI on mortality were investigated.In total, 566 patients with CSD were included in the analyses. A total of 152 patients received the diagnosis of CDI by Toxin A/B EIA, toxigenic culture, or nucleic acid amplification test. Factors associated with CDI included low albumin (adjusted odds ratio (aOR): 1.56; 95% confidence interval (CI): 1.03-2.34) and length of hospital stay before stool collection18 days (aOR: 1.73; 95% CI: 1.09-2.75). CDI was associated with an increased mortality on univariate analysis (OR: 1.6, 95% CI: 1.0-2.6) but was not associated with an increased risk of mortality on multivariable analysis.Risk factors for CDI in Japan were similar to those identified in the USA and Europe. However, CDI was not associated with an increased risk of mortality in this population of patients with CSD.

We have been developing a conduction cooled 3T HTS(REBCO)-MRI system. The prototype system of 100 mm DSV (diameter spherical volume) was designed and made.The MRI image of a mouse fetus (25 mm long) was obtained successfully with 2.9 T under a power supply driven operation. However, the HTS(ReBCO)-MRI magnet still had a considerable magnetic field fluctuation for clear MRI imaging due to a long-lasting attenuation of screening current induced on the superconducting tapes. To solve this problem, we introduced a power supply system consists of a exciter power supply and a small trimming current supply, which compensates the magnetic field deviation due to the screening current, in parallel. We measured and evaluated the magnetic field stability of a commercial 3T-LTS-MRI magnet under both the power supply driven operation with proposed system and the persistent current operation. The availability of the power supply system was discussed using an experimental HTS-MRI system.

Radioactive iodine, one of the radionuclides released in the nuclear power plant accident on 11 March 2011, was detected in purified water at water purification plants (WPPs). However, information about removal of radioactive materials in actual water purification process was limited. Therefore, we investigated the removal of radioactive materials (iodine and cesium) immediately after the detection. It is found that non-radioactive iodine in water could be removed by the combined use of pre-chlorination and powdered activated carbon (PAC) treatment. The same result was also obtained in terms of radioactive iodine. Removal of non-radioactive iodine in WPPs was also investigated. Approximately, 60% of iodine was removed by combination of pre-chlorination (0.5–1.0 mg/L) and PAC (15–30 mg/L) in coagulation and sedimentation processes. In water purification process, cesium was mostly removed by coagulation and sedimentation; hence, radioactive cesium was not detected in purified water. It was confirmed...

Population structure and demographic history of the Japanese Spanish mackerel Scomberomorus niphonius a highly piscivorous and migratory marine fish, were assessed using mitochondrial DNA control region sequences (n = 720) and microsatellite genotypes at five loci (n = 1331) for samples collected on Japanese coasts from 2001 to 2010. The population structure was panmictic and the haplotype and allele frequencies were temporally stable even during the recent recovery process. Demographic expansion was strongly supported throughout the Pleistocene, suggesting that the oscillating glacial and interglacial climate conditions in the Pleistocene had no substantial impact on the demographic history of S. niphonius.

35Cl NQR and XRD have been investigated on the solid solution Cs2[Ag(I) x Au(I)1−x Cl2][Au(III)Cl4] which yielded NQR lines by annealing. The crystal forms a tetragonal cell in the whole range of x. However, a change of the modification occurs at x≈0.85. The increase of c and decrease of a were observed in the range of 0.0≤x≤0.7.

A commercial serodiagnostic kit for diagnosing pulmonary disease due to Mycobacterium avium complex (MAC-PD) was developed and launched in Japan in 2011.To evaluate the performance of this kit in routine clinical settings.In this retrospective single-centre study, data on serum levels of anti-glycopeptidolipid (GPL) core IgA antibody (U/ml) measured using the kit were analysed in patients diagnosed with MAC-PD according to American Thoracic Society criteria, in those with pulmonary tuberculosis (PTB) or pulmonary M. kansasii disease and in healthy volunteers.The anti-GPL-core IgA antibody levels of serum were significantly higher (P0.0001) in patients with MAC-PD (n = 485) than in those with PTB (n = 133) or pulmonary M. kansasii disease (n = 23) or in healthy subjects (n = 265). When the cut-off level was set at 0.7 U/ml, the sensitivity and specificity were respectively 78.6% and 96.9%. Higher antibody levels were observed in patients with greater extent of disease on chest computed tomography (P0.0001).The serodiagnostic kit revealed good sensitivity and specificity. The antibody levels may reflect disease activity. Additional work is needed to determine whether the diagnostic assay could be used in conjunction with current diagnostic criteria to improve the diagnosis of MAC-PD.

This report describes the case of a 76 year old man who suffered from febrile ulceronecrotic Mucha-Habermann disease (FUMHD). Despite this patient's typical clinical and histological findings, the fulminating course led to death. Polymerase chain reaction (PCR) analysis of the skin lesions showed that the infiltrating cells were monoclonal in origin and were from an aberrant clone. FUMHD is a very rare, febrile variant type of pityriasis lichenoides et varioliformis acuta, and is characterised by necrotic cutaneous ulcerations associated with high fever and systemic manifestations. Including this present case, only 18 cases of FUMHD have been reported. FUMHD can occur in both adults and children, although there are several differences between the manifestations of the disease in the two groups. One major difference is prognosis: all cases resulting in fatality are of the adult type, whereas no fatal cases have been reported among children. The aberrant clone detected by PCR may be responsible for host responses, resulting in the severe symptoms observed in this disorder.

Pigmented (PE) and nonpigmented epithelial (NPE) cells were carefully dissected from the ciliary processes of rabbit eyes and maintained in vitro. Both epithelial cultured cells showed hexagonal morphology by light microscopy; abundant granules containing pigment could be seen in PE cells while no pigment was seen in the NPE cells. S-100 protein exists in both cultured cells and was used as a marker for the in vitro confirmation of these cells. Collagen type III was used as a negative marker to rule out stromal cell contamination in tissue cultures. PE cells attached and proliferated well in 20% fetal bovine serum (FBS) in minimum essential media (MEM). Greater proliferation was attained when PE cells were exposed to 20% vitreous extract in addition to 20% FBS-MEM. Media conditioned by PE cells did not significantly stimulate PE cells. NPE cells showed greatest proliferation in 20% FBS-MEM with either 20% concentration of media conditioned by PE cells or when exposed to 20% concentration of vitreous extract. Rabbit vitreous may contain factor(s) which stimulate the proliferation of both cultured cells, and conditioned media from the PE cells may contain factor(s) which stimulate NPE cell proliferation.

The nucleotide sequences of the NP and M genes of human parainfluenza type 1 virus (HPIV-1) were determined. The NP gene was 1677 nucleotides long excluding polyadenylic acid. The NP gene contained a single large open reading frame (ORF), which encoded a polypeptide of 524 amino acids with a calculated molecular weight of 57,736. The M gene 1173 nucleotides long excluding the poly(A) tract and the sequence also contained a single large ORF which encoded a polypeptide of 348 amino acid with a molecular weight of 38,445, which was inconsistent with 28 kDa previously determined by SDS-PAGE. We aligned the deduced HPIV-1 NP and M protein sequences with 12 and 13 other paramyxoviruses, respectively, suggesting that a common tertiary structure was found in the NPs or Ms of HPIV-1, Sendai virus (SV), HPIV-3 and BPIV-3 and that other common structure was also maintained in these proteins of HPIV-2, SV 41 and 5, MuV, HPIV-4. Phylogenetic trees were constructed for the NP and M proteins of all the paramyxoviruses of which nucleotide sequences had been previously reported. Paramyxoviruses could be subdivided into two groups, i.e., PIV-1 group and PIV-2 group; the former group is composed of HPIV-1, SV, HPIV-3 and BPIV-3, and the latter group consists of HPIV-2, SV 41, SV 5, MuV, HPIV-4 A and HPIV-4 B.

An immunohistochemical analysis using antibodies to cytokeratin, epithelial membrane antigen, alpha-1-antitrypsin, alpha-1-antichymotrypsin and factor XIIIa was performed in four cases of malignant fibrous histiocytoma and five cases of sarcomatoid carcinoma in the urinary tract. All cases of malignant fibrous histiocytoma showed positive staining for factor XIIIa, alpha-1-antitrypsin and alpha-1-antichymotrypsin. No case was positive for factor XIIIa, but one case with sarcomatoid carcinoma stained positive for alpha-1-antitrypsin and alpha-1-antichymotrypsin. All cases showed positive staining for cytokeratin and 4 cases with sarcomatoid carcinoma were positive for epithelial membrane antigen, but no cases with malignant fibrous histiocytoma were positive. Immunohistochemical analysis would thus help to distinguish malignant fibrous histiocytoma from sarcomatoid carcinoma of the urinary tract.

Nonpigmented epithelial (NPE) and pigmented epithelial (PE) cells were carefully dissected from both human and rabbit ciliary processes and have been maintained in vitro and partially characterized by morphology and immunocytochemical techniques using polyclonal and monoclonal antibodies against S-100 proteins, collagen type I and type III. The tissue distribution of these proteins was studied in formalin fixed deparaffinized tissue sections of human and rabbit eyes by immunoperoxidase staining techniques. Both NPE and PE cell lines from human and rabbit showed hexagonal morphology by light microscopy; distinct granules containing pigment could be visualized in the PE cell lines, but not in the NPE cells. Antibodies against S-100 proteins stained NPE layer intensely and PE layer slightly in the human tissue sections. The staining was less intense in rabbit tissues than human tissues. The ciliary body stroma was positive for collagen type III and negative for collagen type I or S-100.

Recently, the wearable computing technologies keep empowering a variety of fashion items. Each wearable fashion item is attractive and providing quite new possibilities in terms of new computing devices. However, the conceptual model that supports fashionable coordination among those items is not proposed yet. To address this problem, we present a coordination model for wearable fashion by utilizing the conceptual model of analog synthesizers. We show some behavior styles and these description method with schematic block diagram. A set of prototyping clothing based on the model is also presented for estimations.

This paper proposes a new recurrent neural network and the learning algorithm using time series and steady-state characteristics of nonlinear dynamic systems. Recurrent neural networks are often trained using only time series of systems, but sometimes other information about the system to learn can be obtained. Nonlinear steady-state characteristics of systems are important information to improve performance of recurrent neural networks. Furthermore, this paper shows the computational results to verify the performance of the new recurrent neural network and the learning algorithm.

Compounds that inhibit protein kinases are currently undergoing clinical evaluation for the treatment of a variety of malignancies. The kinase inhibitors flavopiridol and 7 hydroxy-staurosporine (UCN-01) were examined for their effects on B-cell chronic lymphocytic leukemia (B-CLL) cells in vitro (n = 49). Flavopiridol and UCN-01 induced concentration-dependent apoptosis of most B-CLL samples tested, with greater than 50% cell killing occurring at concentrations of less than 1 mcmol/L, and with flavopiridol displaying more potent activity than UCN-01. Flavopiridol (0.1 mcmol/L) and UCN-01 (1 mcmol/L) also induced striking decreases in the levels of the antiapoptosis proteins Mcl-1, X-linked inhibitor of apoptosis (XIAP), and BAG-1 in nearly all cases of B-CLL and of Bcl-2 in approximately half of B-CLL specimens evaluated. In contrast, expression of the proapoptotic proteins Bax and Bak was not significantly influenced by these kinase inhibitors. Flavopiridol-induced decreases in the levels of antiapoptosis proteins Mcl-1 and XIAP preceded apoptosis and were not substantially affected by the addition of caspase inhibitors to cultures. In contrast, UCN-01-stimulated decreases in antiapoptosis proteins were slower, occurred concurrently with apoptosis, and were partially prevented by caspase inhibitors. The findings suggest that flavopiridol and UCN-01 induce apoptosis of B-CLL cells through different mechanisms. The potent apoptotic activities of flavopiridol and UCN-01 against cultured B-CLL cells suggest that they may be effective as single agents in the treatment of B-CLL or for sensitizing B-CLL cells to conventional cytotoxic drugs. (Blood. 2000;96:393-397)

Expression of several inhibitor of apoptosis proteins (IAPs) was investigated in the National Cancer Institute panel of 60 human tumor cell lines, and the expression and prognostic significance of one of these, XIAP, was evaluated in 78 previously untreated patients with acute myelogenous leukemia (AML). XIAP and cIAP1 were expressed in most cancer lines analyzed, with substantial variability in their relative levels. In contrast, NAIP mRNA was not detectable, and cIAP2 was found at the mRNA and protein levels in only 34 (56%) and 5 (8%) of the 60 tumor cell lines analyzed, respectively. Interestingly, XIAP, cIAP1, and cIAP2 mRNA levels did not correlate with protein levels in the tumor lines, indicating posttranscriptional regulation of expression. High levels of XIAP protein in tumor cell lines were unexpectedly correlated with sensitivity to some anticancer drugs, particularly cytarabine and other nucleosides, whereas higher levels of cIAP1 protein levels were associated with resistance to several anticancer drugs. The relevance of XIAP to in vivo responses to cytarabine was explored in AML, making correlations with patient outcome (n = 78). Patients with lower levels of XIAP protein had significantly longer survival (median, 133 versus 52.5 weeks; P = 0.05) and a tendency toward longer remission duration (median, 87 versus 52.5 weeks; P = 0.13) than those with higher levels of XIAP. Altogether, these findings show that IAPs are widely but differentially expressed in human cancers and leukemias and suggest that higher XIAP protein levels may have adverse prognostic significance for patients with AML.

BAG-1 is a multifunctional protein that blocks apoptosis and interacts with several types of proteins, including Bcl-2 family proteins, the kinase Raf-1, certain tyrosine kinase growth factor receptors, and steroid hormone receptors, possibly by virtue of its ability to regulate the Hsp70/Hsc70 family of molecular chaperones. Two major forms of the human and mouse BAG-1 proteins were detected by immunoblotting. The longer human and mouse BAG-1 proteins (BAG-1L) appear to arise through translation initiation at noncanonical CTG codons located upstream of and in-frame with the usual ATG codon used for production of the originally described BAG-1 protein. Immunoblotting experiments using normal tissues revealed that BAG-1L is far more restricted in its expression and is present at lower levels than the more prevalent BAG-1 protein. Human but not mouse tissues also produce small amounts of an additional isoform of BAG-1 of intermediate size (BAG-1M) that probably arises through translation initiation at yet another site involving an ATG codon. All three isoforms of human BAG-1 (BAG-1, BAG-1M, and BAG-1L) retained the ability to bind Hsc70. Subcellular fractionation and immunofluorescence confocal microscopy studies indicated that BAG-1L often resides in the nucleus, consistent with the presence of a nuclear localization sequence in the NH2-terminal unique domain of this protein. In immunohistochemical assays, BAG-1 immunoreactivity was detected in a wide variety of types of cells in normal adult tissues and was localized to either cytosol, nucleus, or both, depending on the particular type of cell. In some cases, cytosolic BAG-1 immunostaining was clearly associated with organelles resembling mitochondria, consistent with the reported interaction of BAG-1 with Bcl-2 and related proteins. Furthermore, experiments using a green fluorescence protein (GFP)-BAG-1 fusion protein demonstrated that overexpression of Bcl-2 in cultured cells can cause intracellular redistribution of GFP-BAG-1, producing a membranous pattern typical of Bcl-2 family proteins. The BAG-1 protein was found at high levels in several types of human tumor cell lines among the 67 tested, particularly leukemias, breast, prostate, and colon cancers. In contrast to normal tissues, which only rarely expressed BAG-1L, tumor cell lines commonly contained BAG-1L protein, including most prostate, breast, and leukemia cell lines, suggesting that a change in BAG-1 mRNA translation frequently accompanies malignant transformation.

The patient was a 48-year-old woman. In January 1995, she noted swelling in the left parotid gland, and saw an otorhinolaryngologist. Needle biopsy showed small cell carcinoma, and she was subsequently admitted to our hospital. Chest radiography revealed a tumor shadow in the hilus of the right lung. Bronchial biopsy revealed small cell carcinoma of the lung (T 4 N 3 M 1, stage IV). Chemotherapy, with a CDDP-VP-16 regimen, achieved no response. She later developed bitemporal hemianopsia and abducens nerve palsy. Brain MRI revealed metastasis in the pituitary gland. Chemotherapy and radiotherapy were efficacious for only a few months. She also developed pain and numbness in the left leg, attributable to intramedullary metastasis (L 1/2, L 4/5) shown on MRI. It is extremely rare for a metastasis to the parotid gland to be the initial clinical manifestation of a small cell lung cancer which later develops widespread metastases to the pituitary gland and lumbar spinal cord.

Immunohistochemical analysis of the apoptosis-effector protease CPP32 (Caspase-3) in normal lymph nodes, tonsils, and nodes affected with reactive hyperplasia (n = 22) showed strong immunoreactivity in the apoptosis-prone germinal center B-lymphocytes of secondary follicles, but little or no reactivity in the surrounding long-lived mantle zone lymphocytes. Immunoblot analysis of fluorescence-activated cell sorted germinal center and mantle zone B cells supported the immunohistochemical results. In 22 of 27 (81%) follicular small cleaved cell non-Hodgkin's B-cell lymphomas, the CPP32-immunopositive germinal center lymphocytes were replaced by CPP32-negative tumor cells. In contrast, the large cell component of follicular mixed cells (FMs) and follicular large cell lymphomas (FLCLs) was strongly CPP32 immunopositive in 12 of 17 (71%) and in 8 of 14 (57%) cases, respectively, whereas the residual small-cleaved cells were poorly stained for CPP32 in all FLCLs and in 12 of 17 (71%) FMs, suggesting that an upregulation of CPP32 immunoreactivity occurred during progression. Similarly, cytosolic immunostaining for CPP32 was present in 10 of 12 (83%) diffuse large cell lymphomas (DLCLs) and 2 of 3 diffuse mixed B-cell lymphomas (DMs). Immunopositivity for CPP32 was also found in the majority of other types of non-Hodgkin's lymphomas studied. Plasmacytomas were CPP32 immunonegative in 4 of 12 (33%) cases, in contrast to normal plasma cells, which uniformly contained intense CPP32 immunoreactivity, implying downregulation of CPP32 in a subset of these malignancies. All 12 peripheral blood B-cell chronic lymphocyte leukemia specimens examined were CPP32 immunopositive, whereas 3 of 3 small lymphocytic lymphomas were CPP32 negative, suggesting that CPP32 expression may vary depending on the tissue compartment in which these neoplastic B cells reside. The results show dynamic regulation of CPP32 expression in normal and malignant lymphocytes.

Allelic loss at nonrandom chromosomal sites is thought to mark the position of tumor suppressor genes involved in the pathogenesis and progression of human malignancies. Solid tumors in particular have been found to harbor multiple genetic changes resulting in loss of function mutations. Tumor suppressor genes have also been found to be involved in the progression of lymphoid tumors. Previous reports have suggested the involvement of a tumor suppressor gene located on the long arm of chromosome 13, between the retinoblastoma (RB) and D13S25 loci, in the pathogenesis and or progression of more than 40% of B-cell chronic lymphocytic leukemia (B-CLL), a common lymphoid malignancy whose molecular etiology remains largely unknown. In the present study, we report the construction and characterization of a YAC contig spanning a region of approximately 3 cM between the RB gene and the D13S31 locus. We also screened 60 paired normal/tumor B-CLL samples for allelic loss on chromosome 13 with nine microsatellite markers located between RB and D13S25. This analysis has allowed us to narrow the smallest region of loss to a segment of 550 kb located between the 206XF12 and D13S25 markers.

An 80-year-old man was admitted to our hospital with complaints of left-sided chest pain and swelling. A chest roentgenogram revealed a left chest-wall mass. Examination of a biopsy specimen of the mass led to the diagnosis of Non-Hodgkin's lymphoma of diffuse mixed-cell type. Immunohistological examination revealed that it was B-cell type. The chest-wall mass was markedly smaller after radiation therapy, and the patient had a complete remission. About 1 year and 10 months later, Non-Hodgkin's lymphoma developed in the right femur. Combination chemotherapy (cyclophosphamide, adriamycin, vincristine, and prednisolone) and radiation therapy were given. The patient went into a complete remission, and is alive at the time of this writing, 2 years and 6 months after he was first treated. Almost all malignant lymphomas of the chest wall arise in cases of chronic tuberculous pyothorax or tuberculous pleuritis, but in this case there was no preceding disease. When the chest roentgenogram shows a chest-wall mass, we must consider the possibility of malignant lymphoma, even if there is no preceding disease.

Lymphoid cells and small intestinal epithelial (SIE) cells are among the most radiosensitive in the body. The factors that account for the differential sensitivity to gamma-radiation among different tissue-types remain poorly understood, but can only partly be explained by differences in rates of cell proliferation. Here we demonstrate that exposure of mice to 800 cGy of gamma-radiation results in rapid elevations in the levels of the Bax protein, a pro-apoptotic member of the Bcl-2 protein family, in lymphoid cells and SIEs. gamma-Radiation-induced increase in Bax protein were evident within 2 h and persisted for at least 24 h, as determined by immunoblotting and immunohistochemical assays. Increases in Bax were followed by massive apoptosis in lymphoid organs and in the small intestinal crypts, as determined by morphological criteria and in situ end-labeling of fragmented nuclear DNA by terminal deoxynucleotidyl transferase (TUNEL method). Radiation did not induce elevations in Bax or apoptosis in radioresistant tissues such as heart, skeletal muscle, brain, kidney, liver, lung, vascular smooth muscle and connective tissue. The findings suggest that Bax may be one of the mediators of radiation-induced apoptosis in vivo.

Oxidative coupling of methane(OCM) over various natural calcium compounds was investigated. Some kinds of shells were found to be good catalysts for OCM. The study of surface properties of the catalysts by XPS revealed that a high performance of the shell catalysts was due to not only morphology but also high concentration of minor components such as sodium on the surface of the catalysts.

To investigate the characteristics of adrenoceptors of the proximal urethra in female rabbits, we performed the in vitro isovolumetric urethral pressure study and isometric study with three parts of muscle strips of the proximal urethra (whole layers, inner layers and outer layers). Both alpha-1 and alpha-2 agonist caused dose dependent response in in vitro isovolumetric pressure study as well as in vivo study. However, the response of alpha-2 agonist in in vitro was small in magnitude compared to in vivo study, suggesting the influence of permeability of drugs from serosa to mucosa in in vitro whole urethra study. The response of the strips of inner layers to alpha-1 agonist is almost the same as that of outer layers and whole layers, while the response of inner layers to alpha-2 agonist is almost twice as that of other layers. These findings are suggestive of predominance of alpha-2 adrenoceptors mediating contractions in the inner layers of the proximal urethra in female rabbits. Alpha-2 adrenoceptors which probably are distributed in mucosal and submucosal layers may have an important role in the mechanism of urinary continence in female rabbits.

We performed combined uroflowmetry and external sphincter electromyography in 108 patients (87 males and 21 females); 16 serving as normal controls, 49 with benign prostatic hyperplasia, 16 with overactive type of neurogenic bladder, 27 with underactive type of neurogenic bladder. Urinary flow pattern was classified into 5 categories. All of urinary flow patterns of normal controls showed pattern A which had the form of a bell. EMG showed complete suppression during micturition followed by a few intermittent increases in amplitude. Urinary flow patterns of BPH patients were distributed widely from pattern A to E. Intermittent increases in EMG amplitude during a late phase of micturition were characteristic in patients with BPH. We must pay attention to sex differences to evaluate urinary flow pattern because it could be influenced by abdominal pressure such as stressing or Créde maneuver in female patients with an underactive type of neurogenic bladder.

Evidence has been found suggesting that long-term therapy with topical antiglaucoma medications may decrease the success of glaucoma filtering surgery. To investigate this question further, the antiproliferative effects of the preservative benzalkonium chloride and three pure and commercially available beta-adrenergic antagonist preparations (timolol, betaxolol, and levobunolol) were studied on tissue cultures of human Tenon's capsule fibroblasts. Each drug preparation was tested on three different cell lines. Fibroblast growth was measured with tritiated thymidine uptake and hexosaminidase assays. Trypan blue uptake was used to assess cell viability microscopically. The commercially available preparations containing benzalkonium chloride and those of betaxolol and levobunolol without the preservative had similar inhibitory doses for 50% of cells. The timolol preparation without preservative was significantly less toxic than its commercially available one. The three tested beta-adrenergic blockers did not stimulate fibroblast proliferation directly in this in vitro model. Even when the cultures were washed free of the drugs, growth continued to be suppressed, suggesting that the inhibition was not reversible. An increase in fibroblasts and inflammatory cells after long-term antiglaucoma medical therapy thus may be caused not by a direct stimulation of cell proliferation but by chronic inflammation from the irritating effects of antiglaucoma medications and/or their preservatives.

To determine the relationship between cardiovascular complications of estrogen therapy and fibrinolysis, fibrinolysis parameters plasminogen, urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor-1 (PAI-1), were assessed in 12 prostatic cancer patients before and 6 weeks after the onset of estrogen therapy. The levels of plasminogen, u-PA, and PAI-1 in the patients treated with the estrogen therapy were significantly higher than those in the patients before the therapy. The t-PA level in the patients during the therapy was significantly lower than that before the treatment. Cardiovascular complications were found in two patients (16.7%) during estrogen therapy. In the two patients, marked elevation of PAI-1 and decreased level of t-PA were observed during the therapy. These results indicate that cardiovascular complications of estrogen therapy in patients with prostatic cancer may be related to hypofibrinolysis resulting from changes of PAI-1 and t-PA.

Collagen deposition is largely responsible for scar information, an undesirable outcome of glaucoma filtering surgery. Because interferon-gamma (IFN-gamma) has been reported to inhibit collagen synthesis in a variety of cells, the authors examined its effect on collagen synthesis by Tenon's capsule fibroblasts cultured from three patients. In addition, the authors studied the effects of IFN-gamma on fibroblast proliferation. IFN-gamma inhibited collagen synthesis at dosages of 1-10,000 U/ml. The average median effective dose (ED50) was 6.60 U/ml. In contrast, IFN-gamma had no significant effect on cell proliferation in dosages of 0.001-10,000 U/ml. These results suggest that IFN-gamma may be a useful agent for modifying the wound healing response, particularly after glaucoma filtering surgery.

5-Fluorouracil (5-FU) has effectively inhibited fibroblast proliferation to prevent scar formation and bleb failure after glaucoma filtering surgery. To identify more potent but less toxic antiproliferative drugs, the authors studied cell attachment and proliferation of 5-FU metabolites: 5-fluorouridine (FUR), 5-fluorodeoxyuridine (FUdR), 5-fluorouridine-5'-monophosphate (FUMP), and 5-fluorodeoxyuridine-5'-monophosphate (FdUMP) on human Tenon's fibroblasts in vitro.

Scar formation and fibrosis are the most common causes of ultimate glaucoma filtration surgery failure. The authors used Coulter counter, hexosaminidase, and 3H-thymidine assays to evaluate the effects of mithramycin, mitomycin, daunorubicin, and bleomycin on human subconjunctival fibroblast growth in tissue culture. Fifty percent inhibitory doses (ID50) for each drug on different days of incubation with fibroblasts were calculated with these three assays. Among these four drugs only daunorubicin exerted a significant effect on cellular attachment, showing cytotoxicity at 4 x 10(-6) M. However, all drugs had antiproliferative effects. Daunorubicin was the most potent drug in the concentration range studied. Mithramycin had a narrow therapeutic range in tissue culture. Cellular antiproliferative responses to mitomycin and bleomycin were more gradual. These drugs have potential applications in the prevention of glaucoma filtration surgery failure.

The in vitro cellular inhibitory effects of two pyrimidine antimetabolites, 5-fluorouracil (5-FU) and cytarabine (ara-C), on the attachment and proliferation of human Tenon's capsule fibroblasts after 1, 2, 3, 5, 7, and 10 days of growth were measured with a Coulter counter, a colorimetric method using the endogenous enzyme hexosaminidase, and 3H-thymidine uptake. Neither 5-FU nor ara-C affected cell attachment. The 50% inhibitory dose (ID50) for 5-FU, as measured by the Coulter counter and hexosaminidase assay, was 0.2 and 0.4 micrograms/ml, respectively, at day 5 and decreased to 0.01 and 0.10 micrograms/ml, respectively, on later days. The ID50 for ara-C as measured by the Coulter counter and hexosaminidase assay was 0.01 and 0.1 micrograms/ml at day 3 and remained constant over time. Much lower ID50s were measured by thymidine uptake for both drugs. These findings may indicate that 5-FU has a delayed effect on cellular proliferation due to conversion into more active metabolites. The ara-C has a direct and constant inhibitory effect on cellular proliferation and is ten times more potent than 5-FU as an antiproliferative drug. Thus ara-C may have clinical utility in preventing failure of glaucoma filtering surgery.

This study demonstrated the importance of the methods used in determining the lymphoma cell colony stimulating activity of factors derived from lymphoma cells. The in vitro colony formation in a semisolid matrix of the AKR mouse lymphoma cell line, SL 12, and three cloned derivatives, SL 12.1, SL 12.3, and SL 12.4, was studied. We show that the use of soft agar or methylcellulose as a semisolid matrix results in colony formation by the lymphoma cells only in the presence of serum. The addition of conditioned medium (CM) from lymphoma cells growing in serum-free medium does not stimulate colony growth. However, when purified agarose is used, colonies grow in a dose-dependent manner in the absence of serum and in the presence of CM. These results indicate that the type of semisolid matrix used can influence results in studies of this nature. Purified agarose provides the best environment when colony formation by lymphoma cells is used to measure the presence of growth factors in test-conditioned media.

The theoretical analysis of a penny-shaped geothermal reservoir in the earth's crust subject to linear tectonic stress gradients has been made on the basis of the three dimensional theory of elasticity. The condition for stability of a reservoir requires K/sub 1/ < K /SUB c/ , where K/sub 1/ and K /SUB c/ are, respectively, the stress intensity factor for the opening mode and the fracture toughness of the surrounding rock. From this condition the upper critical pressure being necessary for the reservoir stability is obtained and is shown graphically.

Autonomic receptor density can be modulated by alterations in neuronal activity over a relatively short period of time (hours). The current study investigates whether increased in vivo stimulation of urinary bladder smooth muscle can alter muscarinic receptor density and response to muscarinic stimulation. A high degree of reflex stimulation of the urinary bladder (rabbits) was initiated by stricture of the external urethra. Intravesical pressure and intra-abdominal pressure were monitored continuously over a 4-hr time period. At the end of the 4-hr period, the rabbits were sacrificed and isolated strips of bladder body were either mounted in isolated smooth muscle baths for contractile studies or frozen and stored in liquid nitrogen for muscarinic receptor analysis. These studies demonstrated that over 4 hr of urethral stricture there was a significant reduction in muscarinic receptor density from a Bmax of 34 +/- 3.4 fmol/mg of protein in control bladder strips to 22 +/- 2.4 fmol/mg of protein in the experimental group. In association with the decreased muscarinic receptor density, there was a significant and selective decrease in the contractile response to muscarinic stimulation. Similar to the in vivo studies, repetitive field stimulation of in vitro strips resulted in a significant decrease in muscarinic receptor density and a significant and selective decrease in the contractile response to muscarinic stimulation. The results from these studies indicate that muscarinic receptor density, and response to muscarinic stimulation, can be modulated over a relatively short period of time by alterations in the level of neuronal stimulation.

The role of tumor cell membrane gangliosides in tumor formation was probed using a series of cloned murine AKR lymphoma cell lines. Tumor formation was directly related to high expression and shedding of membrane gangliosides. In vivo, as little as 1 pmol of purified total gangliosides of highly tumorigenic cells, injected intradermally with poorly tumorigenic cells (which lacked and did not shed gangliosides), markedly increased the tumorigenicity of these cells in syngeneic normal mice. Thus, gangliosides shed by tumor cells are a previously unrecognized, extremely potent enhancer of tumor formation in vivo.

Transferrin has been considered to be an essential requirement for hematopoietic cell proliferation in culture. We have isolated two cloned lymphoma cell lines, SL 12.1 and SL 12.4, which grow and adapt in serum-free medium without added transferrin. Antibody to the transferrin receptor blocks the growth of these cells. We have also demonstrated that transferrin-free conditioned medium from the cells will compete with transferrin for binding. Furthermore, conditioned medium from SL 12.1 and SL 12.4 cells induces and supports exponential growth of a transferrin-dependent lymphoma cell line, SL 12. We conclude that these two transferrin-independent cloned lines produce transferrin-like activity which plays a crucial role for cell proliferation.