Four new solvates using ethanol, isopropyl alcohol, this website acetonitrile, and toluene vapor and one polymorph using water vapor were discovered. Their solid state characterizations were conducted using powder X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis, and Fourier transform infrared spectroscopy. A variety of isostructural solvates and a polymorph of TDF was produced by an organic vapor sorption method, showing varying physicochemical properties. This study

demonstrates an alternative crystallization method to obtain isostructural solvates. (C) 2013 Elsevier B.V. All rights reserved.”
“Epirus is a rural area of North-Western Greece. We reviewed data from 4 GDC 973 hospitals for 4.975 patients who underwent prostate biopsy in Epirus in the twelve year period from 1999 to 2010. Two six -year periods were compared

(1999-2004 and 2004-2010). All cases of prostate cancer confirmed by biopsy were recorded and age-standardized incidence rates per 100,000 males were calculated. We also recorded the clinical stage for patients diagnosed in our hospital and correlated this with PSA and Gleason scores. Percentage of positive prostate biopsies was also calculated. There were a total of 1714 new cases during 1999-2010 and the mean annual age-adjusted incidence was 34/100.000. The mean incidences during 1999-2004 and 2005-2010 were 26/100,000 and 42/100,000, respectively. The mean age at diagnosis was 74. The most common Gleason score was 6 and the BAY 73-4506 prevalent clinical stage was T2. Median PSA at diagnosis was 10.8 ng/ml. There was a significant difference between stage cT4 and all other stages regarding PSA value (p=0.000). A positive correlation was found between Gleason score and PSA (p=0.013). These results are in accordance with the incidence rise recorded in neighboring countries of South-East Europe. However we should keep in mind the risk of overdiagnosis and the detection of low-risk cancers that would not have caused morbidity or death during a man’s lifetime anyway.”
“We investigate the microscopic structure and

density fluctuations of complex coacervates of flexible polyelectrolytes using scattering of neutrons, X-rays, and light. Poly(acrylic acid) and poly(N,N-dimethylaminoethyl methacrylate) offer a well-defined model system that allows for selective labeling and systematic variation of the strength of the attractive electrostatic interactions. Two neutron scattering experiments have been carried out: (i) we use deuterated polymeric tracers in a complex coacervate with an overall neutron scattering length density that is matched to that of the solvent, to probe the conformation of single polymer chains in the complex coacervates, and (ii) we measure complex coacervates in which all polymer chains of one type are deuterated, to probe their overall structure.

The importance of amplitude on pain relief has not

been established. The current study aims to: a) investigate the importance Of amplitude as part of the treatment dose. b) To explore the extent of any pain reliving effects seen following mobilisations.\n\nThe study employed a randomised, single blind, within-subjects repeated measure design. Thirty asymptomatic subjects participated. click here The subjects completed three experimental conditions on three separate occasions. The conditions were: large amplitude of oscillations (forces between 50 and 200 N), small amplitude of oscillations (150 N-200 N) and quasi-static (maintained at 200 N). Each condition involved a 3×1 minute central PA mobilisation at a frequency of 1.5 Hz on the lumbar spine. Pressure pain thresholds (PPT) were measured immediately before and after each intervention at 4 different sites. The sites were chosen to determine the extent of the hypoalgesic response.\n\nResults demonstrated a significant increase in PPT following lumbar mobilisations (p = 0.013)

at all measured sites. However, no significant difference was found between amplitude conditions (p = 0.864). This study suggests that in asymptomatic subjects a systemic hypoalgesic response is caused by lumbar mobilisation regardless Of amplitude. (C) 2009 Elsevier Ltd. All rights reserved.”
“MR spectroscopy is a noninvasive technique that allows the detection of several naturally occurring selleck chemicals llc compounds (metabolites) from well-defined regions of interest within the human brain. Alzheimer disease, a progressive neurodegenerative disorder, is the most common cause of dementia in the elderly. During the past 20 years, multiple studies have been performed on MR spectroscopy in patients with both mild cognitive impairment and Alzheimer

disease. Generally, MR spectroscopy studies have found decreased N-acetylaspartate and increased myo-inositol in both patients with mild cognitive impairment and Alzheimer disease, with greater changes in Alzheimer disease than in mild cognitive impairment. This review summarizes the information content of proton brain MR spectroscopy and its related technical aspects, as well as applications ofMRspectroscopy to mild cognitive impairment and Alzheimer disease. While MR spectroscopy may have some value in the differential Tipifarnib cell line diagnosis of dementias and assessing prognosis, more likely its role in the near future will be predominantly as a tool for monitoring disease response or progression in treatment trials. More work is needed to evaluate the role of MR spectroscopy as a biomarker in Alzheimer disease and its relationship to other imaging modalities.”
“A career in science is shaped by many factors, one of the most important being our tastes in research. These typically form early and are shaped by subsequent successes and failures. My tastes run to microscopes, chemistry, and spatial organization of cytoplasm.

g., 3,3′,4′,5-trans-tetrahydroxystilbene (piceatannol), 3,3′,5,5′-trans-tetrahydroxystilbene (3,3′,5,5′-THS) and 3,3′,4′,5,5′-trans-pentahydroxystilbene (3,3′,4′,5,5′-PHS). All these compounds were cytotoxic Prexasertib to growth-arrested C6 cells, with EC(50)-values between 20 and 85 mu M. A higher cytotoxic potency in proliferating cells indicated a specific cytostatic activity of resveratrol and 3,3′,4′,5,5′-PHS. All hydroxystilbenes studied inhibited cellular radical generation induced by cumene hydroperoxide (CH P). The rank order of antioxidant potency was resveratrol >

piceatannol > 3,3′,5,5′-THS>3,3′,4′,5,5′-PHS. However, only resveratrol and piceatannol inhibited cellular radical generation at lower than cytotoxic concentrations. At subcytotoxic concentrations only piceatannol was able to protect the cells from damage caused by CHP. Taken together, these results show that neither the cytotoxic or cytostatic activities of hydroxystilbenes nor their cytoprotective

and antioxidant activities in living cells can be predicted from their antioxidant and prooxidant activity, respectively, in cell-free systems. (C) 2009 Elsevier Ireland Ltd. All rights reserved.”
“In Alzheimer disease (AD), the perturbation of the endoplasmic reticulum (ER) calcium (Ca2+) homeostasis has been linked to presenilins, the catalytic core in gamma-secretase complexes CH5183284 cleaving

the amyloid precursor protein (APP), thereby generating amyloid-beta (A beta) peptides. Here we investigate whether APP contributes to ER Ca2+ homeostasis and whether ER Ca2+ could in turn influence A beta production. We show that overexpression of wild-type human APP (APP(695)), or APP harboring the Swedish Galunisertib supplier double mutation (APP(swe)) triggers increased ryanodine receptor (RyR) expression and enhances RyR-mediated ER Ca2+ release in SH-SY5Y neuroblastoma cells and in APPswe-expressing (Tg2576) mice. Interestingly, dantrolene-induced lowering of RyR-mediated Ca2+ release leads to the reduction of both intracellular and extracellular A beta load in neuroblastoma cells as well as in primary cultured neurons derived from Tg2576 mice. This A beta reduction can be accounted for by decreased Thr-668-dependent APP phosphorylation and beta- and gamma-secretases activities. Importantly, dantrolene diminishes A beta load, reduces A beta-related histological lesions, and slows down learning and memory deficits in Tg2576 mice. Overall, our data document a key role of RyR in A beta production and learning and memory performances, and delineate RyR-mediated control of Ca2+ homeostasis as a physiological paradigm that could be targeted for innovative therapeutic approaches.

The diagnosis of DCRV should be considered in the young patient with an elevated right ventricular systolic pressure and intracavitary gradient. Echocardiography and cardiac MRI are the principal diagnostic tools for the assessment of DCRV. This entity is often misdiagnosed PI3K inhibitor as pulmonary

hypertension in the young patient, and can often go overlooked and untreated for many years. Definitive therapy involves surgical resection of the muscle bundle. This can often be curative and if done in a timely fashion, may prevent right ventricular remodeling. We describe the unique diagnostic dilemma, the course and management of a young adult with DCRV during pregnancy.”
“Prunus mira Koehne ex Sargent (syn. Persica mira (Koehne) Kov. et Kostina), native to China, is an excellent fruit tree due to its high ecological and economical value. However, there is limited knowledge on the genetic information of P. mira. In this study, the genetic relationships of 83 P. mira accessions from five populations were assessed using amplified fragment

length polymorphism (AFLP). The results showed that AFLP was a powerful tool to detect levels of genetic diversity of natural populations in P. mira. The similarity coefficient between accessions ranged from 0.12 to 0.76, with an average 0.57. 83 accessions were clustered into two major clusters at similarity coefficient of 0.225. The highest values of N (e), H and I occurred in ML population. Most of the genetic Selleckchem ALK inhibitor variations occur

within population. There is no close relationship between geographic distance and genetic distance. At the same time, ex situ conservation needs to be established for P. mira.”
“Ulcerative colitis (UC) is characterized by chronic inflammation of the colon. During inflammation, NF-kappa B is increased in colonic epithelial cells and in immune cells, leading to increases in proinflammatory cytokines. These events then increase DNA methyltransferases (DNMTs), which silence a subset of tumor suppressor genes by promoter methylation. Negative regulators of the Wnt pathway are frequently methylated in UC, leading to dysregulation of the pathway and, BVD-523 nmr potentially, to colorectal cancer. We determined if black raspberries (BRBs) influence promoter methylation of suppressors in the Wnt pathway in dextran sodium sulfate (DSS)-induced UC. C57BL/6J mice received 1% DSS and were fed either control or 5% BRB diets. Mice were euthanized on days 7, 14 and 28, and their colons, spleen and bone marrow were collected. Berries reduced ulceration at day 28. This was accompanied by decreased staining of macrophages and neutrophils and decreased NF-kappa B p65 nuclear localization in the colon at all time points. At day 7, BRBs demethylated the promoter of dkk3, leading to its increased messenger RNA (mRNA) expression in colon, spleen and bone marrow.

25). In APP, mortality risk of pups was primarily influenced by prey availability as lower beaver density in western APP negatively PARP activation influenced survival and increased starvation risk. Genetic ancestry only influenced pup mortality risk outside of APP with eastern wolf x coyote hybrids surviving poorly relative to other Canis types in WMU49. Poor survival of pups in western APP reduces dispersal from the protected area and decreases the likelihood of expansion of this genetically distinct eastern wolf population beyond APP. Our results advance understanding of wolf and coyote demography as no previous studies have investigated genetic and environmental factors influencing mortality

CDK inhibitor of wolf and coyote pups smaller than 4-5 months of age with telemetry data. (C) 2013 Elsevier Ltd. All rights reserved.”
“In the present paper, in view of the variational approach, we discuss the nonlinear eigen-value problems for p(x)-Laplacian-like operators, originated from a capillary phenomenon. Under some suitable conditions, we prove the existence of nontrivial solutions of the system for every parameter lambda bigger than 0. (C) 2014 Elsevier Ltd. All rights reserved.”
“CXCR4 is a

coreceptor of HIV-1 infection in host cells. Through a photocrosslinking study to identify receptors involved in internalization of oligoarginine cell-penetrating peptides (CPPs), we found that CXCR4 serves as

a receptor that stimulates macropinocytic uptake of the arginine Immunology & Inflammation inhibitor 12-mer peptide (R12) but not of the 8-mer. We also found that stimulating CXCR4 with its intrinsic ligands, stromal cell-derived factor 1 alpha and HIV-1 envelope glycoprotein 120, induced macropinocytosis. R12 had activity to prevent viral infection for HIV-1(IIIB), a subtype of HIV-1 that uses CXCR4 as a coreceptor for entry into susceptible cells, whereas the addition of a macropinocytosis inhibitor, dimethylamiloride, resulted in enhancement of viral infection. The present study shows that CXCR4 triggers macropinocytosis, which may have implications for the cellular uptake of oligoarginine CPPs and internalization of HIV.”
“Respiratory substrates and adenine nucleotides cross the mitochondrial outer membrane through the voltage-dependent anion channel (VDAC), comprising three isoforms – VDAC1, 2, and 3. We characterized the role of individual isoforms in mitochondrial metabolism by HepG2 human hepatoma cells using siRNA. With VDAC3 to the greatest extent, all VDAC isoforms contributed to the maintenance of mitochondrial membrane potential, but only VDAC3 knockdown decreased ATP, ADP, NAD(P)H, and mitochondrial redox state. Cells expressing predominantly VDAC3 were least sensitive to depolarization induced by increased free tubulin. In planar lipid bilayers, free tubulin inhibited VDAC1 and VDAC2 but not VDAC3.

The suitability

of routinely measured quantities for the prediction of tablet quality was tested. Results showed that model-based expert systems based 5-Fluoracil DNA Damage inhibitor on the contemporary routinely measured quantities can significantly improve the trial-and-error procedures: however, they cannot completely replace them. The modelling results also suggest that in cases where it is not possible to obtain sufficient number of measurements to uniquely identify the model, it is beneficial to use several modelling techniques to identify the quality of model prediction. (c) 2009 Elsevier B.V. All rights reserved.”
“Local delivery of growth factors (GFs) can accelerate regeneration of injured tissue, but for many medical applications, injectable GF delivery systems are required for clinical success. Viscoelastic, injectable aggregates of micrometer-sized hydrogel particles made of multiarmed polyethylene glycol (starPEG) and heparin were prepared and tested for site-specific paracrine stimulation of tissue regeneration. Heparin was used as it binds, protects and releases numerous GFs. Hydrogel based delivery

of basic fibroblast growth factor (bFGF) and murine epidermal growth factor (EGF) was monitored utilizing enzyme-linked immunosorbent assay (ELISA). bFGF was released slowly because of its high affinity to the heparin while the significantly higher release of the non-specific binding EGF was controlled by diffusion only. To investigate GF delivery in vivo, a hydrogel loaded with murine EGF or bFGF AZD9291 was injected P505-15 nmr subcapsularly into the left kidney of mice with experimental acute kidney injury caused by glycerol induced rhabdomyolysis. Visual examination confirmed sustained stability of the injected gel aggregates during the timescale of the experiment. The number of proliferating kidney tubular epithelial cells was quantified both in the injected kidney and the non-injected contralateral kidney. bFGF

delivery from hydrogels induced a significant increase in cell proliferation in the injected kidney, although small effects were also seen in the non-injected kidney due to a systemic effect. EGF delivery strongly increased cell proliferation for both kidneys, but also showed a local effect on the injected kidney. The hydrogel without loaded GFs was used as a control and showed no increase in cell proliferation. Our results suggest that this novel starPEG-heparin hydrogel system can be an effective approach to deliver GFs locally. (C) 2013 Elsevier B. V. All rights reserved.”
“The alternative pathway of the complement cascade plays a role in the pathogenesis of dense deposit disease (DDD). Deficiency of complement factor H and mutations in CFH associate with the development of DDD, but it is unknown whether allelic variants in other complement genes also associate with this disease.

Lineage tracing using a tamoxifen-inducible Cre expressed from the Tcf21 locus demonstrated that the majority of Tcf21-expressing epicardial cells are committed to the cardiac fibroblast lineage prior to initiation of epicardial epithelial-to-mesenchymal transition (EMT). Furthermore, Tcf21 null hearts fail to form cardiac fibroblasts, and lineage tracing of the null cells showed their inability to undergo EMT. This is the first report of a transcription factor

essential for the development of cardiac fibroblasts. We demonstrate a unique role for Tcf21 in multipotent epicardial progenitors, prior to the process of EMT that is essential for cardiac fibroblast development.”
“A nucleosides containing block copolymer, poly(polyethylene glycol methyl ether methacrylate)-block-poly(5′-O-methacryloyluridine) (PPEGMEMA(30)-b-PMAU(80)) was self-assembled in aqueous medium and GSK621 price cross-linked via RAFT polymerization at 60 degrees C to afford core-cross-linked NVP-BSK805 mouse micelles exhibiting a PPEGMEMA corona and a polynucleotide core. A disulfide cross-linking agent, bis(2-methacryloyloxyethyl)disulfide, was employed to cross-link the structure via the RAFT process resulting in core-shell nanoparticles, which can degrade under reductive conditions.

The resulting core-cross-linked micelles readily hydrolyzed into free block copolymers in the presence of dithiothreitol (DTT) in less than 1 h, depending on the concentration of the reducing agent and the amount of cross-linker in the micelle. A small fraction of permanently cross-linked micelle was found as the result of conventional chain transfer to disulfide containing compounds. A model drug, vitamin B-2, was loaded into the micelle. The loading capacity increased with increasing cross-linking degree. The amount of drug released reached 60-70% after 7 h in the presence of DTT (0.65 mM), while the cross-linked micelle in the absence of dithiothreitol shows only a delayed drug release. Cytotoxicity

tests confirmed the biocompatibility of the polymers and the residues after reduction.”
“Goal: Selleck Nocodazole The goal of this study is to evaluate the safety and efficacy of Small intestinal release mesalamine (SIRM) for symptom relief in refractory celiac disease (RCD).\n\nBackground: Therapeutic options for the RCD are inadequate and treatment with corticosteroids and immunosuppressants is limited by side effects. SIRM has been shown to have local antiinflammatory action and excellent tolerability.\n\nStudy: We reviewed records of the RCD patients who received SIRM in an open-label therapeutic trial. Data included demo-graphics, disease characteristics, dose and duration of SIRM therapy, and response. Response was categorized as complete if there was complete resolution of symptoms, partial if there was at least 50% improvement, and nonresponsive if there was less than 50% improvement.

training on one labeled stack and testing ASP1517 on one from a different subject with similar skin type. Initial results demonstrate the detectability of the DE) in both scenarios with epidermis/dermis misclassification rates smaller than 10% and average distance from the expert labeled boundaries around 8.5 mu m. (C) 2011 Society of Photo-Optical Instrumentation

Engineers (SPIE). [DOI: 10.1117/1.3549740]“
“Background: Dovitinib (TKI-258) is a receptor tyrosine kinase (RTK) inhibitor targeting fibroblast growth factor receptor (FGFR) and further related RTKs. TKI-258 is under investigation as anticancer drug for the treatment of various cancers including bladder cancer with aberrant RTK signaling. Here, we analyzed the responses of ten human bladder cancer cell lines towards TKI-258 treatment in relation to the epithelial mesenchymal transition (EMT) status AS1842856 cost of the cells.\n\nMethods: Expression of epithelial marker E-cadherin as well as mesenchymal markers N-cadherin and vimentin was determined by quantitative RT-PCR and Western-blot in RNA and protein extracts from the cultured cell lines. The cell responses were analyzed upon addition

of TKI-258 by viability/proliferation (XTT assay) and colony formation assay for measurement of cell contact independent growth.\n\nResults: The investigated bladder cancer cell lines turned out to display quite different EMT patterns as indicated by the abundance of E-cadherin or N-cadherin and vimentin. Protein and mRNA levels of the respective components strongly correlated. Based on E-cadherin and N-cadherin mRNA levels that were expressed approximately mutual exclusively, an EMT-score was calculated for each cell line. A high EMT-score indicated mesenchymal-like cells and a low EMT-score epithelial-like cells. Then, we determined the IC50 values for TKI-258 by dose response curves (0-12 mu M TKI-258) in XTT assays for each cell line. Also, we measured the clonogenic survival fraction after adding TKI-258 (1 mu M) by colony formation assay. We observed significant correlations between EMT-score

and IC50 values (r = 0.637, p = 0.0474) and between EMT-score and clonogenic survival fraction (r = 0.635, p = 0.0483) as analyzed by linear regression analyses.\n\nConclusions: In sum, we demonstrated that the EMT status based on E-cadherin buy PF-03084014 and N-cadherin mRNA levels may be useful to predict responses towards TKI-258 treatment in bladder cancer.”
“We report a joint analysis of positron annihilation lifetime spectroscopy (PALS), dielectric spectroscopy (BDS), and nuclear magnetic resonance (NMR) on cis-trans-1,4-poly(butadiene) (c-t-1, 4-PBD). Phenomenological analysis of the orthopositronium lifetime tau(3) – T dependence by linear fitting reveals four characteristic PALS temperatures: T G T-b1(G) = 0.63T(g)(PALS), T-g(PALS), T-b1(L) = 1.22T(g)(PALS), and T-b2(L) = 1.52T(g)(PALS).

12 [95% CI 1.00, 1.26] per additional medication) and the measure of basic activities of daily living Barthel Index (RR = 0.94 [95% CI 0.88, 0.99] per increase) were independently associated with the use of hospital days.\n\nConclusion: Exposure to DBI medications was associated with a greater use of hospital days, but a cumulative dose-response relationship between DBI and hospitalization was not observed. The number of regularly used medications and functioning BEZ235 in the basic activities of daily living predicted hospital

care utilization.”
“Background/Aims: The frequency of mixed hepatitis B virus (HBV) genotypes in chronic HBV (CHB) and genotype changes during natural disease evolution and as a result of antiviral

therapy were investigated.\n\nMethods: Serum samples from 103 CHB patients were included in a cross-sectional study. Longitudinal study of HBV genotypes was performed in 22 patients, 17 of them under antiviral therapy (lamivudine and/or adefovir). HBV genotyping was done by the INNO-LiPA HBV assay.\n\nResults: Genotypes observed in the cross-sectional study: A 32% of cases, D 42%, C 2%, F 2%, and mixed genotypes 22% (mainly A/D, followed by A/G). Genotype G was found in 7% of patients, always combined with other genotypes. In the longitudinal study, genotype changes were observed only in treated patients (9 cases). Genotype A strains were positively selected in 6 of them, mainly as mixed AID. In 6 patients, this website selection coincided with a decrease in HBV-DNA levels.\n\nConclusions: A high frequency of mixed HBV genotypes was observed in our setting. Selection of genotype A strains during treatment is likely an indication that sensitivity to therapy differs between genotypes A and D. The absence of changes in untreated patients suggests that HBV genotype is stable without external factors. (C) 2008 European Association for the Study of the Liver. Published by

Elsevier B.V. All rights reserved.”
“The yellow fever virus (YFV), the first proven human-pathogenic virus, although isolated in 1927, is still GSK1120212 research buy a major public health problem, especially in West Africa where it causes outbreaks every year. Nevertheless, little is known about its genetic diversity and evolutionary dynamics, mainly due to a limited number of genomic sequences from wild virus isolates. In this study, we analyzed the phylogenetic relationships of 24 full-length genomes from YFV strains isolated between 1973 and 2005 in a sylvatic context of West Africa, including 14 isolates that had previously not been sequenced. By this, we confirmed genetic variability within one genotype by the identification of various YF lineages circulating in West Africa. Further analyses of the biological properties of these lineages revealed differential growth behavior in human liver and insect cells, correlating with the source of isolation and suggesting host adaptation.

A J-shaped dose response, in the form of a significant decreased response at some low dose followed by a significant increased

response at a higher dose, was found in only four datasets from three experiments. Three of these datasets involved the same control animals and two also shared dosed animals; the J shape in the fourth dataset appeared to be the result of an outlier within an otherwise monotonic dose response. A meta-analysis was conducted to determine whether AZD7762 there was an excess of dose groups with decreases in tumor response below that in controls at doses below no-observed-effect levels (NOELs) in individual datasets. Because the probability of a decreased response is generally not equal to the probability of an increased response even in the null case, the meta-analysis focused on comparing the number of statistically significant diminished responses to the number expected, assuming no dose effect below the NOEL. Only 54 dose groups out of the total of 2579 in the database had doses

below the dataset-specific NOEL and that satisfied an a priori criterion for sufficient power to detect a reduced response. Among these 54, a liberal criterion for defining a significant decreases identified 15 such decreases, versus 54 x 0.2 = 10.8 expected. The excess in significant reductions was accounted for almost entirely by the excess from neutron experiments (10 observed, 6.2 expected). Nine of these 10 dose groups involved only 2 distinct control groups, MEK162 MAPK inhibitor and 2 pairs from the 10 even shared dosed animals. Given this high degree of overlap, this small excess did not appear remarkable, although the overlap prevented a formal statistical SNX-5422 order analysis. A comprehensive post hoc evaluation using a range of NOEL definitions and alternative ways of restricting the data entering the analysis did not produce materially different results. A second meta-analysis found that,

in every possible low dose range ([0, d] for every dose, d) of each of the radiation types, the number of dose groups with significantly increased tumorigenic responses was either close to or exceeded the number showing significantly reduced responses. This meta-analysis was considered to be the more definitive one. Not only did it take dose into account by looking for consistent evidence of hormesis throughout defined low-dose ranges, it was also potentially less susceptible to limitations in experimental protocols that would cause individual animals to respond in a non-independent fashion. Overall, this study found little evidence in a comprehensive animal radiation database to support the hormesis hypothesis. However, the ability of the database to detect a hormetic effect was limited both by the small number of dose groups with doses below the range where positive effects have been found in epidemiological studies (<= 0.