Substantial differences in the analyte profiles were notable, with the group demonstrating the highest level of periodontitis showing elevated levels of IL-6, IL-8 and LBP and significantly decreased levels of PGE2 and BPI. By the time of delivery, and following ligation of teeth in four quadrants, all animals had a CIPD >20 (not periodontally healthy). Again, the most diseased animals provided a profile of serum analytes that was distinctive from animals expressing primarily gingival inflammation,

with a lower level of destructive disease. These data suggested that the variation in naturally occurring periodontal find more inflammation and disease in the female baboons was reflected by patterns of systemic inflammation. Moreover, those animals that responded more robustly to the infection burden accompanying ligation generally

demonstrated a unique profile of mediator levels. As we have observed previously, these findings are consistent with a subset of these non-human primates that show an increased susceptibility to dysregulated local responses eliciting greater disease and allowing a more substantial challenge to the systemic inflammatory apparatus. These outcomes would also suggest that animals with a more effective adaptive immune response to the microbial challenge would demonstrate less disease, as we have reported previously [46,55], and less systemic challenge with lower serum inflammatory responses. Examination of the relationships between the inflammatory mediators and antibody in serum showed that elevated or decreased antibody specificities were coincident RG-7388 nmr with levels of selected mediators. However, identification of a particular pattern of antibodies that best described the systemic inflammatory response profiles was somewhat complex. Generally, the acute phase reactants were delineated by

unique patterns of antibody responses that were observed at specific time-points during the study. The chemokines IL-8 and MCP-1 demonstrated some similarity in the patterns of antibody correlations, particularly at baseline Dynein and mid-pregnancy. IL-6 levels were best described by distinctive antibody specificities during the protocol. However, of the 20 antibody specificities that were evaluated, levels of F. nucleatum, P. gingivalis, A. actinomycetemcomitans and C. rectus showed some consistency in contributing to relationships with the range of inflammatory mediators analysed. However, within the model system, a pattern of the serum analytes provided some insight into describing the expression of disease. We observed a clear association of IL-6, IL-8 and LBP levels across disease expression and throughout pregnancy. When broken down further, we observed that these relationships were related primarily to the characteristics of the disease expression in the individual animal, and generally related less to the stage of pregnancy at which the sample was obtained.

23,111 Danger and stress selleck kinase inhibitor signals following allergen encounter or parasite invasion can invoke danger-associated molecular patterns (DAMPs) such as ATP.113–115 ATP, in addition to TLR signalling, can potently activate the inflammasome leading to IL-1β processing, which has been shown by several groups to enhance Th2 effector responses.89,116–118 Interestingly, blood dwelling schistosomes posses ATP-catabolizing enzymes on their tegument surfaces that breakdown ATP to adenosine, potentially interfering with this pathway.119 Following differentiation, Th2 cells are distinguishable from

Th1 cells by more than just cytokine gene activation. For example, Th2 cells lose the ability to sustain calcium flux 120 resulting in reduced tyrosine phosphorylation.121 Th2 cells also have an unconventional synapse, relative to Th1 and naive T cells, and fail to form a ‘bulls-eye’ structure.122 These apparent differences may be because of reduced CD4 and increased CTLA-4 expression, as suggested by others.123 The consequences of these structural selleck chemicals llc differences between Th1 and Th2 cells are unclear. Unlike IFN-γ, which is secreted directionally in the immunological synapse, IL-4 can be secreted multi-directionally influencing many surrounding cells.124,125 Whether this is a result of altered

synapse formation or not has not been reported. Also, whether IL-5 and IL-13 are indiscriminately secreted multi-directionally within the reactive lymph node has not been reported. The precise activation

signals received by differentiated Th cells, stimulating their effector function are rather vague. For example it may not be desirable for a Th2 cell, or Th1, Th17 or Th9 cell, to release their payload Selleckchem Lonafarnib of potent cytokines, beyond polarizing IL-4, in the case of Th2, within the T-cell zones of lymphoid tissue. Therefore, restricted re-activation via peptide-loaded MHC-II-expressing cells or other activating signals at the site of infection, allergy or action must take place. What these additional signals are is surprisingly unclear. Following Th2 differentiation, chromatin remodelling at conserved non-coding sequence (CNS)-1, DNase I hypersensitivity (DHS) site, CNS-2 and the conserved intron 1 sequence of IL-4 (CIRE) in the il4 locus facilitates rapid cytokine transcription.126–128 Poised in such a state, it may only require a ‘tickle’ to induce translation and secretion of these cytokines. An elegant study by Mohrs et al.,129 using a dual reporter system to identify transcription and secretion of IL-4, discovered that although IL-4 was transcribed in lymphoid and non-lymphoid tissue, secretion was only observed in non-lymphoid tissue upon antigen encounter. This study is in slight contradiction to a recent paper from the same group identifying the widespread influence of IL-4 in the reactive lymph node.

1b. The bars represent the mean BrdU+CD19+ absolute cell numbers and the standard error of the mean represent quintiplicate cultures. The differences in cell numbers among the different co-cultures are not statistically significant (two-way analysis of variance). Fig. S4. A representative flow cytometric analysis that underlies the data shown in Fig. 1c,d is shown. The magenta-coloured values represent the frequency of the specific cell populations as a percentage of the parental flow cytometric Ceritinib gate. Fig. S5. The isotype controls used to establish the acquisition gates for the flow cytometric

analysis of the co-cultures described in Fig. 2a are shown. Fig. S6. Confirmation that Aldefluor+ cells reside inside the CD11c+ cell population in control dendritic cells (cDC) and immunosuppressive DC (iDC) generated from peripheral blood mononuclear cells (PBMC) of two unrelated healthy adult individuals. The magenta coloured values represent the frequency of Aldefluor+ cells inside the CD11c+ gate. Aldefluor mea fluorescence intensity (MFI) is shown in magenta

colour at the bottom of the specific histograms. “
“Recent research has shown that (i) Toll-like receptor (TLR) agonists drive hematopoietic stem and progenitor cells buy BMS-777607 (HSPCs) to proliferate and differentiate along the myeloid lineage in vitro, and (ii) direct TLR-mediated stimulation of HSPCs also promotes macrophage differentiation in vivo following infection. These

new insights demonstrate that TLR signaling in HSPCs, in addition to other TLR-dependent mechanisms, can contribute to HSPC expansion and myeloid differentiation O-methylated flavonoid after infection. Evidence is, therefore, mounting that direct TLR-induced programming of hematopoiesis plays a key role in host defense by rapidly replenishing the innate immune system with the cells needed to deal with pathogens. Throughout life, leukocytes arise from a common ancestor in the mammalian BM, the hematopoietic stem cell (HSC), which is functionally defined by its durable capacity for self-renewal and ability to produce all types of blood cells (Fig. 1, reviewed in [1, 2]). During homeostasis, the process of HSC self-renewal, as well as the production of lineage-committed progenitors, is tightly controlled to maintain daily blood cell production. Many cytokines, cell–cell interactions, and transcription factors “fine-tune” the proliferation of hematopoietic stem and progenitor cells (HSPCs) and their differentiation into mature myeloid and lymphoid cells (reviewed in [3]). Upon infection, or during other forms of immunological stress, there is an increased demand for leukocytes to assist in combating the infection, to replace cells killed by invading microbes or consumed during the immune response, and to increase immune surveillance. The adaptive immune system meets this demand by clonal expansion of T and B cells.

Hemolymph (100 µL) was collected from both treated and control groups and centrifuged at 800 g for 5 mins (Model GS-15R, Rotor No. F2402; Beckman, Fullerton, CA, USA). After centrifugation, the supernatant was discarded, the hemocytes washed three times with Hank’s buffered salt solution and then stained with NBT solution (0.3%, 100 µL) for 30 mins at 37°C. The staining reaction was terminated by removing the NBT solution and adding absolute methanol. After three washings with 70% methanol, the hemocytes were air dried and 120 µL of 2-M KOH and 140 µL of DMSO added to dissolve cytoplasmic formazan. The optical density of the dissolved formazan was

read at 630 nm. Alkaline and acid phosphatase activities assays were performed according to the methods described by Gestal

et al. [23]. Briefly, ALP and ACP were measured using p-nitro phenyl phosphate CT99021 cell line 16 mM as a standard substrate. Glycine NaOH buffer and sodium acetate buffer were used for ALP and ACP assays, respectively. ABT-737 Mixtures containing 0.2 mL of the substrate and 50 µL of hemolymph were incubated for 30 min at 37°C. Released p-nitrophenol in the resulting supernatants was measured at 410 nm and the amount calculated from the standard curve. One-way ANOVA followed by Tukey’s test was performed to identify significant differences among experimental groups at each sampling time using Statistical Analysis Software (SAS Institute, Cary, NC, USA). For statistically significant differences, an α value of < 0.05 (P < 0.05) was required. Linear regression analysis (comparison

between biochemical and immune variables and salinity of WSSV-challenged hemolymph of F. indicus) was performed to analyze WSSV infection and the influence of each salinity concentration. The unchallenged control F. indicus kept in 25 g/L survived. Mortality began at 24 hrs in the challenged shrimp kept in 5 and 35 g/L. Over all 24–96 hrs, the cumulative mortality of F. indicus maintained in 5 and 35 g/L was significantly higher than that of shrimp kept in 25 and 15 g/L (P < 0.05). At 72 hrs pi, the cumulative mortality of challenged F. indicus maintained in 25 g/L was the lowest among the experimental groups, whereas the cumulative mortality of the challenged F. indicus transferred to 5 g/L was the highest among the four treatments. No mortality was recorded in any of the unchallenged groups during the experimental period. In WSSV challenged animals, mortality increased in parallel with sampling time. For all salinity concentrations except for 25 g/L salinity, the mortality rates ranged from 63.3 ± 3.3% (15 g/L) to 83.3 ± 3.3% (5 g/L). From the start of the experiment (24th hour), animals exposed to 5 g/L salinity had a mortality of 53.3 ± 3.3%. However, animals at 25 g/L showed a comparatively lower mortality rate after infection with WSSV (Table 1). Total hemolymph protein concentration increased significantly at 48 and 72 hrs pi (P < 0.

The effects had never been studied yet on a lung model for large mammals. Our data showed dose-dependent effects of CsA on gas exchanges, but also on pulmonary hemodynamics, and possibly an aggravation of the IRI due to high doses of CsA. These results constitute an important step toward the use of CsA on humans to reduce lung IRI and consequently, primary graft dysfunction. Within a few years, the EVLP technique has become a reference for the evaluation of lung grafts. Its interest has been demonstrated see more on animal

lung preparations, especially on pig [43] and human lungs [12]. This technique can be seen as bench test for lung function, allowing for the assessment of new therapies suppose to limit IRI. Gas exchange capacities and total pulmonary arterial resistance are more commonly studied physiopathological parameters. We also measured other hemodynamics (Pcap,

longitudinal pulmonary resistance) and markers (AFC, RAGE, cytokines, lung permeability) that have showed their pertinence in the evaluation of lung IRI [5, 7]. It has been hypothesized that IRI is mostly related to mitochondrial death as a consequence of MPTP opening. Located in the inner mitochondrial membrane, the MPTP remains unremarkable under normal physiological conditions. Stress can lead to its opening, resulting in the swelling of the matrix due to osmotic forces. It then induces further failure of the mitochondrial outer membrane and the release of the cytosol pro-apoptotic factors [19]. The inhibition of Rapamycin cell line the opening of MPTP is thought to be the main pathway for CsA action. Several in vitro and in vivo animal models showed CsA interests in pre and post-conditioning for the

prevention of IRI on different organs such as heart, kidney, and liver [19, 20, 45, 50]. In humans, CsA administered just before coronary reperfusion (post-conditioning) has been proven to be an efficient way to reduce the size of myocardial infarction [33]. However, few studies have been published on CsA effects on lung IRI. In vitro studies on post hypoxia-reoxygenation injuries showed that alveolar macrophages pretreated by CsA secreted less chemokines than Myosin controls [30]. Moreover, endothelial cells incubated with CsA selectively reduced pro-inflammatory mediator secretion of NFκB and EGR-1 [15]. Nevertheless, some of the pathways involved in IRI can be activated by CsA, such as the metalloproteinase and the TLR [1, 28, 41]. Such insights can explain the increased levels of pro-inflammatory cytokines we measured in our experiments with high doses of CsA (30 μM). In an in vivo ischemic lung model, Krishnadasan et al. showed that rats pre-conditioned with CsA displayed less tissue myeloperoxidase content, leukocyte accumulation, and vascular permeability [25].

falciparum infection, and our observations disclose clear differences associated with progression and regression of malaria tropica. This work was conducted at the Centre Hospitalier Regional (CHR) in Sokodé in the Central Region of Togo. The study was approved by the Comite de Bioethique pour la Recherche en Sante (CBRS) in Togo, and by the Ethikkommission at University Clinics of Tübingen,

Germany. Informed written consent was obtained from all parents for the participation of their children find more in this study. Infants of less than 5 years of age were recruited, and classification of malaria was performed according to previously published criteria [14], with severe malaria (SM) characterized by parasitaemia of higher than 250 000parasites/µl and/or the presence of severe anaemia with haemoglobin concentrations of lower than 5 g/dl. Matched uncomplicated malaria (MM) patients were defined by parasitaemia of lower than 250 000 parasites/µl and haemoglobin concentrations equal to or higher than 5 g/dl and the absence of any signs or symptoms of severe malaria [13]. P. falciparum-exposed infants negative for parasites in thick www.selleckchem.com/products/LY294002.html blood film, and negative in rapid detection test kits for P. falciparum (Paracheck-Pf, Orchid, Biomedical Systems, Goa, India; OptiMAL-IT; Biorad, Marnes la Coquette, France),

were defined as participants with previous malaria episode(s) and the actual absence of illness due to malaria within the last 2 weeks. Blood samples were obtained prior to treatment with anti-malarials and/or anti-pyretics, and immediately following primary diagnosis all P. falciparum-positive infants received anti-malarial and appropriate supportive therapy as required and recommended by the Guidelines for Malaria Treatment indicated by the Ministry of Health in Togo. Infants with MM were treated with Coartem and Artemeter or Artesunate, and for SM, quinine perfusion or injectable Artemeter

were applied as recommended. All hospitalized uncomplicated as well as severe malaria cases were followed until discharge from the hospital paediatric ward. Quantitative enzyme-linked immunosorbent assay (ELISA) was performed with commercially available assays to determine Glutathione peroxidase plasma levels of the cytokines IL-10, IL-13, IL-17F, IL-27, IL-31 and IL-33, as well as of the chemokines MIP3-α/CCL20, monokine induced by gamma interferon (MIG)/CXCL19, 6Ckine/CCL21 and CXCL16 (Duo-Set; R&D Minneapolis, MN, USA). Sample concentrations of each cytokine and chemokine were quantified from standard curves generated with recombinant chemokines/cytokines, and the lower limit for their detection was 50 pg/ml. For data analyses the statistical package jmp version 5·0.1·2 was used. For the cytokine and chemokine analyses, differences between groups were determined after logarithmic transformation to stabilize the variance of data [log (pg/ml + 1)].

HIF-1α(+/+; −/−) mouse embryonic fibroblast cell lines were kindly provided by Dr. Randall Johnson (University of California, San Diego). Cells were cultured in α-modified Eagle medium or in Dulbecco’s Modified Eagle’s medium, both of which were supplemented with 10% heat-inactivated fetal calf serum, in a 5% CO2 humidified atmosphere at 37°C. The oxygen tension in the chamber was either 20% (normoxic) or 1% (hypoxic). Chaetocin and other chemicals were administered to medium 1 hour before normoxic or hypoxic incubation. Male nude mice (BALB/cAnNCrj-n/n) were purchased from Charles CDK inhibitor River Japan (Shin-Yokohama, Japan) and housed in a specific pathogen-free room. Mice

(6 weeks old) were injected subcutaneously in a flask with 5 × 106 viable cancer cells. After tumor volumes reached 100-150 mm3, mice were treated with dimethyl sulfoxide (DMSO), chaetocin (0.25 mg/kg, intraperitoneally [i.p.]), and/or doxorubicin (1 mg/kg, i.p.) once a day. Tumor volumes CHIR-99021 mw were measured with a caliper and calculated using the equation volume = ab2/2, where “a” is the maximal width and “b” is maximal orthogonal width. All procedures were conducted in accordance with the guidelines of the Laboratory Animal Ethics Committee of Seoul National University. All data were analyzed using Microsoft Excel 2007 or

SPSS (v. 10.0) software and results are expressed as means and 95% confidence intervals or standard deviations. The two-sided Mann-Whitney U test was used to compare luciferase activities, vascular endothelial growth factor (VEGF) concentrations, and HIF-1α-positive cell, CD31-positive vessel, and Transferase-Mediated dUTP Nick-End Labeling (TUNEL)-positive cell numbers. Tumor sizes were compared using analysis of variance (ANOVA) followed by Duncan’s multiple range test. Differences were considered significant for P < 0.05. All statistical tests were two-sided. To examine if chaetocin has anticancer activity against

hepatoma, chaetocin was injected for 2 weeks into mice bearing mTOR inhibitor Hepa 1c1c-7 tumors. Tumor growth was significantly retarded after chaetocin treatment (Fig. 1A). Interestingly, chaetocin did not induce massive cell death or deformation (Fig. 1B, upper), suggesting that cytotoxicity does not primarily underlie the anticancer effect. Moreover, neither histological changes nor apoptosis was observed in the livers of mice treated with chaetocin for 2 weeks (Fig. 1B, lower). Because a hypoxic microenvironment and angiogenesis are critical for tumor growth, we assessed HIF-1α expression and vascular density immunohistochemically. In chaetocin-treated tumors, HIF-1α-positive cells and CD31-positive threadlike vessels were noticeably reduced, but TUNEL-positive apoptotic cells increased (Fig. 1B, upper); the results are summarized in Fig. 1C. Furthermore, VEGF mRNA was down-regulated overall in chaetocin-treated tumors (Fig. 1D).

[71-73] Migraineurs have lower interictal pain thresholds than controls, suggestive of abnormal sensory-discriminative processing, and lower pain tolerance thresholds suggestive of abnormal affective responses to pain.[5, 74] CM also has deleterious effects on mood and cognitive abilities. Irritability,

depression, anxiety, difficulty concentrating, and impairments in executive function are common during and between migraine attacks.[75, 76] Consistent with the wide-ranging phenotypic expression of migraine, the findings of this rs-fc study suggest that migraine involves numerous aspects of the pain experience, including affective, sensory-discriminative, FDA-approved Drug Library cell line and cognitive domains. Atypical rs-fc between anterior

insula and pulvinar AZD1208 cost might relate to migraine intolerance to light, the abnormal perception of visual stimuli as painful, and/or visual salience.[77] Because the pulvinar receives inputs from dura-sensitive spinal trigeminal nucleus neurons and from the optic nerves, it is postulated that the pulvinar participates in the integration of visual stimuli with trigeminal nerve-mediated head pain.[23, 78, 79] Pulvinar-mediated integration may help to explain why: (1) 40% of migraineurs have light-triggered migraines; (2) >90% of migraineurs have light hypersensitivity (photophobia) during attacks; (3) headache intensity and photophobia intensity are positively correlated; (4) exposing interictal migraineurs to bright light leads to reduced pain thresholds in trigeminal innervated locations, an effect not detected in controls; (5) painful forehead stimulation in interictal migraineurs, but not controls, leads to decreased visual discomfort thresholds; (6) compared with controls and migraineurs without allodynia, migraineurs

with interictal allodynia have altered cortical visual processing.[80-83] Atypical rs-fc of the anterior insula with middle temporal cortex could relate to migraine intolerance to auditory stimuli and to migraineurs misperception of normally nonpainful auditory stimuli as painful.[7] Auditory stimuli interact with migraine in several ways: (1) 50-75% of migraineurs have noise-triggered migraines; (2) >90% of migraineurs have sound hypersensitivity (phonophobia) Teicoplanin during migraine attacks; (3) headache intensity positively correlates with phonophobia intensity; (4) interictal sound hypersensitivity is reported by ∼75% of migraineurs; (5) sound aversion thresholds are lower in interictal migraineurs compared with controls.[6, 7, 50, 84] Future studies will explore relationships between quantitative measures of light and sound hypersensitivity with functional connectivity strength between affective pain regions with pulvinar and affective pain regions with middle temporal cortex.

When the contrast effect in the tumor was greater or smaller than the range of intensity variability in the parenchyma, the lesion was defined as hyper- or hypo-enhancement. In cases where the contrast check details effect in the tumor was within the range of the intensity variability, the lesion was defined as iso-enhancement. All data were expressed as the mean ± standard deviation (SD), median, or percentage. Continuous variables were analyzed by Student t-test or Mann–Whitney U-test. Categorical variables were analyzed using the Fisher exact test or chi-squared test. The cumulative rates were analyzed by Kaplan–Meier

method, and the multivariate analyses were assessed by Cox regression using the best cut-off value obtained from receiver operating characteristics curves. P-values < 0.05 were considered to be significant. Statistical analysis was performed using the SAS software (version 9.2; SAS Institute, Cary, NC, USA). CEUS was performed in 222 patients with 321 lesions during the study period. However, because follow-up was not performed for 13 patients with 19 lesions, CEUS findings were examined for a total of 209 patients with 302 lesions (Fig. 1). A total of 72 subjects (45 males and 27 females; age 65.0 ± 10.8 years) with 87 PIELs (Tables 1 and 2) met the inclusion and exclusion criteria. The mean lesion diameter was 12.5 mm (SD, 4.2 mm; range

5–26.5). The median observation period was 22.0 months (3.3–53.1). Twenty patients

selleckchem developed HCC lesions during the study period; a single lesion was detected in nine patients, two lesions in two patients, and three or more lesions in nine patients. Diagnosis of HCC was made by CEUS, CT, and MRI in 12, by CT and MRI in four, by CEUS and CT in three, and by CEUS and MRI in one. The mean diameter of HCC at the time of detection/diagnosis was 15.1 ± 4.0 mm (10.0–28.6). The overall cumulative HCC occurrence rates were 7.9% at 1 year, 26.3% at 2 years, and 36.0% at 3 years. A total of 14 patients had developed HCC originating from PIELs, and six patients had HCCs not from PIELs. Although there were three PIELs that showed arterial-phase hyper-enhancement new at the time of detection, their diameter and contrast-enhanced appearance remained unchanged, and they did not progress to HCC during the follow-up periods of 22.2, 23.3, and 30.6 months. Univariate analysis showed that the presence of coexistent HCC (P = 0.001) and alpha-fetoprotein (AFP) > 20 ng/mL (P = 0.002) were significant factors at baseline for HCC occurrence. The overall cumulative HCC occurrence rates were significantly higher in patients with coexistent HCC (n = 29; 11.1% at 1 year, 59.9% at 3 years) than those without coexistent HCC (n = 43; 5.7% at 1 year, 17.3% at 3 years; P = 0.001) (Fig. 2), and in patients with AFP > 20 ng/mL (n = 22; 16.3% at 1 year, 68.

Tolvaptan at 7.5 mg/day Selleckchem PLX4720 was considered the optimal dose in liver cirrhosis patients with hepatic edema who showed

inadequate response to conventional diuretics. LIVER CIRRHOSIS REPRESENTS the end stage of any chronic liver disease.[1] Hepatic edema including ascites and lower limb edema is the most frequently observed complication in the disease, leading to deterioration in quality of life.[2, 3] Therefore, improvement of hepatic edema is an important therapeutic strategy. Spironolactone, an aldosterone antagonist, either alone or in combination with the loop diuretic furosemide is prescribed as the first-line therapy for management of liver cirrhosis patients with persistent ascites.[4] Many patients with ascites do not respond to diuretic therapy or require administration of diuretics at high doses that can cause adverse events including activation of the rennin–angiotensin and sympathetic nervous systems, electrolyte disturbances PLX3397 mouse such as dilutional hyponatremia, and worsening of renal function.[5-8] Thus, the development of

effective drugs other than conventional diuretics is needed for the management of hepatic edema. Because arginine vasopressin V2 receptor antagonists promote electrolyte-free water excretion without disrupting electrolyte balance, they are expected to be clinically useful in the treatment of diseases associated with hyponatremia or fluid retention.[9, 10] Tolvaptan, a novel aquaretic agent, is a non-peptide V2 receptor antagonist.[10-12] almost By inhibiting reabsorption at the renal collecting tubules, tolvaptan increases electrolyte-free urine excretion without increasing electrolyte excretion. In the USA,[13] tolvaptan at 15–60 mg/day has been approved for the treatment of hyponatremia, and in the EU,[14] tolvaptan within the same dosage range has been approved for the treatment of syndrome of inappropriate antidiuretic hormone. In Japan, tolvaptan at 15 mg/day has been approved for the treatment of heart failure-related edema.[15] We initiated a program to obtain the additional indication for the treatment of hepatic edema. Therefore, we conducted

the phase 2 study to determine an optimal dose of tolvaptan. In our previous, preliminary trial, tolvaptan at dose of 15 mg/day and higher exerted sufficient pharmacological response for improvement of hepatic edema including ascites in liver cirrhosis patients who showed resistance to furosemide.[16] The aim of this trial was to determine the optimal dose of tolvaptan in hepatic edema showing inadequate response to conventional diuretics. The results of this trial will be used as the basis for a pivotal trial to be conducted to obtain an additional indication for tolvaptan in Japan. THE PRESENT TRIAL was a randomized, double-blind, placebo-controlled, multicenter trial conducted by Otsuka Pharmaceutical (the study sponsor).