41 [1.131.76] for PRS, 0.68 [0.590.79] for PFS and 0.87 [0.731.04] for overall survival, respectively. In conclusion, thalidomide exposed upfront correlated with shorter PRS that partially compensated BAY 73-4506 Protein Tyrosine Kinase inhibitor for prolonged initially PFS and resulted in no survival benefit when it is given as both induction pre-autologous and maintenance post-autologous stem cell transplantation; shorter PRS was not observed, and survival was improved when it is given only during maintenance phase following autologous stem cell transplantation in the patients with myeloma and who are eligible for transplant. Copyright (c) 2011 John Wiley & Sons, Ltd.”
“An ultra-performance liquid chromatography tandem

mass spectrometry (UPLC-MS/MS) method employing electrospray ionization (ESI) has been developed for the determination of mangiferin in rat plasma using diphenhydramine as the internal standard (IS). Liquid-liquid extraction (LLE) was used for sample preparation and the analysis was achieved with

gradient elution on C(18) reversed phase column. The method was validated over the concentration range 0.02-5.0 mu g/mL for oral administration and 0.4-100 mu g/mL for intravenous administration. The intra-day and inter-day precision of mangiferin expressed as RSD < 15% and the accuracy (RE) did not exceed 15%. This validated method is a novel technique for sample preparation and quantitation, which was successfully Cytoskeletal Signaling inhibitor applied to estimate the bioavailability of mangiferin. (C) 2009 Elsevier B.V. All rights reserved.”
“Background\n\nAdenoidectomy, surgical removal of the adenoids, is a common ENT operation worldwide

in children with otitis AZD1480 nmr media. A systematic review on the effectiveness of adenoidectomy in this specific group has not previously been performed.\n\nObjectives\n\nTo assess the effectiveness of adenoidectomy versus non-surgical management or tympanostomy tubes in children with otitis media.\n\nSearch strategy\n\nWe searched the Cochrane Ear, Nose and Throat Disorders Group Trials Register; the Cochrane Central Register of Controlled Trials (CENTRAL); PubMed; EMBASE; CINAHL; Web of Science; BIOSIS Previews; Cambridge Scientific Abstracts; mRCT and additional sources for published and unpublished trials. The date of the most recent search was 30 March 2009.\n\nSelection criteria\n\nRandomised controlled trials comparing adenoidectomy, with or without tympanostomy tubes, versus non-surgical management or tympanostomy tubes only in children with otitis media. The primary outcome studied was the proportion of time with otitis media with effusion (OME). Secondary outcomes were mean number of episodes, mean number of days per episode and per year, and proportion of children with either acute otitis media (AOM) or otitis media with effusion (OME), as well as mean hearing level.

In this pathway, PINK1 accumulates on defective mitochondria, eliciting the translocation of PARKIN from the cytosol to mediate the clearance of damaged mitochondria via autophagy (mitophagy). Throughout the different stages of mitophagy, post-translational modifications (PTMs) are critical for the regulation of PINK1 and PARKIN activity and function. Indeed, activation and recruitment of PARKIN onto damaged mitochondria involves PINK1-mediated phosphorylation of both PARKIN and Ub. Through a stepwise cascade, PARKIN is converted from an autoinhibited

enzyme into an active phospho-Ub-dependent E3 ligase. Upon activation, PARKIN ubiquitinates itself in concert with many different mitochondrial substrates. The Ub conjugates attached to DAPT inhibitor these substrates can in turn be phosphorylated by PINK1, which triggers further find more cycles of PARKIN recruitment and activation. This feed-forward amplification loop regulates both PARKIN activity and mitophagy. However, the precise steps and sequence of PTMs in this cascade are only now being uncovered. For

instance, the Ub conjugates assembled by PARKIN consist predominantly of noncanonical K6-linked Ub chains. Moreover, these modifications are reversible and can be disassembled by deubiquitinating enzymes (DUBs), including Ub-specific protease 8 (USP8), USP15, and USP30. However, PINK1-mediated phosphorylation of Ub can impede the activity of these DUBs, adding a new layer of complexity to the regulation of PARKIN-mediated mitophagy by PTMs. It is therefore

evident that further insight into how PTMs regulate the PINK1-PARKIN pathway will be critical for our understanding of mitochondrial quality control.”
“Brook trout Salvelinus fontinalis (Mitchill, 1814) chromosomes IWR-1-endo concentration have been analyzed using conventional and molecular cytogenetic techniques enabling characteristics and chromosomal location of heterochromatin, nucleolus organizer regions (NORs), ribosomal RNA-encoding genes and telomeric DNA sequences. The C-banding and chromosome digestion with the restriction endonucleases demonstrated distribution and heterogeneity of the heterochromatin in the brook trout genome. DNA sequences of the ribosomal RNA genes, namely the nucleolus-forming 28S (major) and non-nucleolus-forming 5S (minor) rDNAs, were physically mapped using fluorescence in situ hybridization (FISH) and primed in situ labelling. The minor rDNA locus was located on the subtelo-acrocentric chromosome pair No. 9, whereas the major rDNA loci were dispersed on 14 chromosome pairs, showing a considerable inter-individual variation in the number and location. The major and minor rDNA loci were located at different chromosomes. Multichromosomal location (3-6 sites) of the NORs was demonstrated by silver nitrate (AgNO3) impregnation. All Ag-positive i.e. active NORs corresponded to the GC-rich blocks of heterochromatin.

Primary research into if or how MDT models of care improve outcomes for women with complex pregnancies is urgently needed.”
“Background and purpose:\n\nalpha-Humulene and trans-caryophyllene are plant sesquiterpenes with pronounced anti-inflammatory properties. Here, we evaluated the effects of these compounds in an experimental model of airways allergic inflammation.\n\nExperimental approach:\n\nFemale BALB/c mice, sensitized to and challenged with ovalbumin received daily alpha-humulene GSK3326595 inhibitor or trans-caryophyllene (50 mg center dot kg-1, orally) or alpha-humulene (1 mg center dot

mL-1, by aerosol) as either a preventive (for 22 days) or therapeutic (from the 18th to the 22nd day) treatment. Dexamethasone or budesonide was used as a positive control drug. Inflammation was determined on day 22 post-immunization by leukocyte recruitment, interleukin-5 (IL-5), CCL11, interferon-gamma (IFN-gamma) and leukotriene (LT)B(4) levels in bronchoalveolar lavage fluid (BALF). In addition, transcription factors [nuclear factor kappa B (NF-kappa B), activator protein 1 (AP-1)] and P-selectin in lung tissue were measured by immunohistochemistry and mucus secretion by histochemistry.\n\nKey check details results:\n\nPreventive or therapeutic treatments with alpha-humulene,

but not with trans-caryophyllene, significantly reduced the eosinophil recruitment to the BALF. In addition, alpha-humulene recovery INF-gamma and reduced the IL-5, CCL11 and LTB(4) levels in BALF, as well as

the IL-5 production in mediastinal lymph nodes (in vitro assay). Furthermore, alpha-humulene decreased the NF-kB and the AP-1 activation, the expression of P-selectin and the increased mucus secretion in the lung.\n\nConclusions and implications:\n\nalpha-Humulene, Apoptosis inhibitor given either orally or by aerosol, exhibited marked anti-inflammatory properties in a murine model of airways allergic inflammation, an effect that seemed to be mediated via reduction of inflammatory mediators, adhesion molecule expression and transcription factors activation.\n\nThis article is part of a themed issue on Mediators and Receptors in the Resolution of Inflammation. To view this issue visit http://www3.interscience.wiley.com.qe2a-proxy.mun.ca/journal/121548564/issueyear?year=2009.”
“Genomic imprinting in gametogenesis marks a subset of mammalian genes for parent-of-origin-dependent monoallelic expression in the offspring. In mice, the identification and manipulation of individual imprinted genes has shown that the diverse products of these genes are largely devoted to controlling pre- and postnatal growth. Human syndromes with parental origin effects have been characterized both at the phenotypic and genotypic levels, allowing further elucidation of the function and regulation of imprinted genes. Evidence suggests that a compromised in utero environment influences fetal growth through the modulation of epigenetic states.

sakazakii biogroups. MLVA successfully discriminated among these isolates and compared favorably with pulsed-field gel electrophoresis. The method was relatively fast and easy to perform. The potential value of MLVA as an epidemiological tool is discussed.”
“In the title compound, Selleckchem HIF inhibitor C(28)H(35)N(3)O(4), the piperidine ring adopts a boat conformation while the piperazine ring adopts a chair conformation with an equatorial

orientation of the phenyl groups. The dihedral angle between the mean planes of the benzene rings is 74.14 ( 8)degrees. The molecular conformation is stabilized by a weak intramolecular C-H center dot center dot center dot N interaction and the crystal packing is stabilized by weak intermolecular C-H center dot center dot center dot O interactions.”
“The multi-dimensional quantum hydrodynamic equations for charge transport in ultra-small electronic devices like semiconductors, where quantum effects (like particle tunnelling through potential barriers and built-up in quantum wells) take place, is considered in the present paper, and the recent progress on well-posedness, stability analysis, and small scaling limits are reviewed.”
“Objective To investigate the early and delayed effects of cavernous nerve electrocautery injury (CNEI) in a rat model, with the expectation that this model could be used to test rehabilitation therapies for erectile dysfunction

(ED) after radical prostatectomy (RP). Materials and Methods In all, 30 male Sprague-Dawley rats were randomly divided equally into two groups (15 per group). Caspase inhibitor The control group

received CNs exposure surgery only and the experimental group received bilateral CNEI. At 1, 4 and 16 weeks after surgery (five rats at each time point), the ratio of maximal intracavernosal pressure (ICP) to mean arterial pressure (MAP) was measured in the two groups. Neurofilament expression in the dorsal penile nerves was assessed by immunofluorescent staining and Masson’s trichrome staining was used to assess the smooth muscle to collagen ratio in both groups. Results At the 1-week follow-up, KPT-8602 clinical trial the mean ICP/MAP was significantly lower in the CNEI group compared with the control group, at 9.94% vs 70.06% (P smaller than 0.05). The mean ICP/MAP in the CNEI group was substantially increased at the 4- (35.97%) and 16-week (37.11%) follow-ups compared with the 1-week follow-up (P smaller than 0.05). At all three follow-up time points, the CNEI group had significantly decreased neurofilament staining compared with the control group (P smaller than 0.05). Also, neurofilament expressions in the CNEI group at both 4 and 16 weeks were significantly higher than that at 1 week (P smaller than 0.05), but there was no difference between 4 and 16 weeks (P bigger than 0.05). The smooth muscle to collagen ratio in the CNEI group was significantly lower than in the control group at the 4- and 16-week follow-ups (P smaller than 0.

“
“The objective of this study was to investigate product performance of freeze dried l-arginine/sucrose-based formulations under variation of excipient weight ratios, l-arginine counter ions and https://www.selleckchem.com/products/ganetespib-sta-9090.html formulation pH as a matrix to stabilize a therapeutic monoclonal antibody

(MAb) during freeze drying and shelf life. Protein and placebo formulations were lyophilized at aggressive primary drying conditions and key attributes of the freeze dried solids were correlated to their thermal properties and critical formulation temperature. Stability (physical) during processing and long-term storage of the MAb in different formulations was assessed by SE-HPLC. Thermal properties of the mixtures were greatly affected by the type of l-arginine

counter ion. High glass transition temperatures were achieved by adding multivalent acids, whereas the temperature values significantly decreased in the presence of chloride ions. All mixtures were stable during freeze drying, but storage buy AZD1480 stability varied for the different preparations and counter ions. For l-arginine-based formulations, the protein was most stable in the presence of chloride ion, showing no obvious correlation to estimated global mobility of the glass. Besides drying behavior and thermal properties of the freeze dried solids, the counter ion of l-arginine

must be considered relevant for protein shelf life stability. (c) 2015 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:2345-2358, 2015″
“Our clinical experience has suggested that the presently recommended patch-test concentration (1.0%) for formaldehyde in the baseline series might be too low. Therefore, consecutively patch-tested dermatitis patients were tested simultaneously with formaldehyde 1.0% and 2.0% (w/v) in aqua. Formaldehyde 1.0% and 2.0% were applied with a micro-pipette (15 mu l) to filter paper discs in Finn Chambers (0.30 mg/cm(2) and 0.60 mg/cm(2), respectively). A total of 1397 patients with dermatitis were patch-tested. see more In all, 68 (4.9%) patients reacted positively to formaldehyde; 37 reacted only to 2.0%, 29 reacted to both concentrations, and 2 reacted only to 1.0%. Significantly more patients were thus diagnosed with contact allergy to formaldehyde 2.0% compared with 1.0% (p<0.001). We detected 0.1%, 0.4%, and 29.6% irritant reactions to 1.0%, 2.0%, and 3.0% formaldehyde, respectively. We conclude that, with an optimized patch-test technique, doubling the dose per area detects significantly more contact allergies to formaldehyde, but an even higher test concentration causes too many irritant reactions to be usable.

So far, few studies have evaluated the roles of NKT cells

in the pathogenesis this website of aplastic anemia (AA), an autoimmune disease. In this study, we investigated the quantitative and qualitative changes in NKT cells in bone marrow (BM) mononuclear cells of AA patients in response to in vitro stimulation with alpha-galactosylceramide. Compared to healthy controls, BM from AA patients had reduced fraction of NKT cells, which possessed a decreased potential to expand in vitro in response to alpha-galactosylceramide stimulation, producing more IFN gamma+NKT1 cells. In the presence of rhG-CSF, the expansion capacity of NKT cells stimulated by alpha-galactosylceramide was significantly reduced in both AA and control

groups, with the majority of the activated NKT cells expressing intracellular IL-4, and the fractions of IFN gamma + NKT cells were significantly reduced. In summary, our results indicate that polarization of NKT cells towards the NKT2 sub-population occurs after co-stimulation with alpha-galactosylceramide and rhG-CSF in AA. Copyright (C) 2008 S. Karger AG, Basel.”
“Carcinogenesis is a complex process involving both genetic and epigenetic mechanisms. The cellular molecular epigenetic machinery, including histone modifications, is associated with changes Selleck MI-503 in gene expression induced by exposure to environmental agents. In this paper, we systematically reviewed publications regarding the effects of xenobiotic stressors, mainly heavy metal exposure, on specific histone modifications. We included a total of 18 publications describing

the effect of environmental stressors on histone structure modifications. We then constructed an interaction map to visualize the effect of environmental exposure(s) on specific histone modifications. In the studies Selleck WH-4-023 we considered, a total of 20 modifications were reported, of which H3Me3K4 and H3Me2K9 were the most frequently studied histone modifications. These modifications were affected mostly by heavy metals and ethanol exposure. Based on the interaction map, we explored the molecular mechanisms mediating the histone modifications induced by environmental stressors in the respective selected studies. This resulted in the identification of seven target proteins and two families of proteins mediating the effects of environmental stressors on histone modifications. This review contributes to the understanding of environmental exposure and its possible effects on cancer risk by inducing changes in histone modifications and hence gene expression.

\n\nTo study the role of specific

chemical compounds in affecting the herbivore community, we used cultivated Brassica oleracea. The cultivars differ qualitatively in glucosinolate profile, i.e., foliar composition of different glucosinolate compounds, and only a little in total concentration of glucosinolates, the secondary metabolites specific for the Brassicaceae family. In field and laboratory experiments, we tested whether individual compounds explained differences in herbivore community composition, and whether herbivores with a similar degree of host plant specialization responded in a similar way to variation in glucosinolate profiles.\n\nIn the field B. oleracea cultivars differed widely in species richness and composition of the herbivore community, selleck kinase inhibitor as well as in the density of insects they harbored. Plants with high concentrations of the short side chain alkenyl glucosinolate, glucoiberin, harbored low herbivore diversity. Higher biodiversity was found when plants had glucosinolate profiles containing high concentrations of glucosinolates with elongated side chains, which are biosynthetically linked to glucoiberin. Although glucosinolates are known to have differential effects on generalist and specialist herbivores,

all herbivore species exhibited similar responses to the intraspecific variation in foliar glucosinolate profiles of the learn more B. oleracea selleckchem cultivars. This

observation is supported by the correspondence between oviposition preferences of the specialist herbivore Pieris rapae and the generalist Mamestra brassicae in the field and the laboratory, using the same cultivars, and may be due to the relatively low concentrations of glucosinolates in cultivars. Our results show that variation in the concentration of short side-chain glucosinolates affects the composition of the herbivore community associated with brassicaceous plants.”
“Eastern wolves (Canis lycaon) are a species of special concern federally in Canada that have hybridized extensively with coyotes (Canis la trans) and gray wolves (Canis lupus) in and adjacent to Algonquin Provincial Park (APP), Ontario. A comprehensive understanding of eastern wolf demography and wolf-coyote hybridization dynamics is needed for informed management, yet no studies have investigated the influence of genetic ancestry and environmental heterogeneity on survival of wolf, coyote, and hybrid pups. We used telemetry, genetic, and environmental data to model and estimate genotype-specific survival and cause-specific mortality of Canis pups (n = 159) in eastern and western APP (2002-2011) and the adjacent Wildlife Management Unit 49 (WMU49; 2008-2011). Pup survival was higher in eastern APP ((s) over cap= 0.75) and WMU49 ( smaller than (s)over cap = 0.

Mild to moderate asymptomatic hyperglycemia is observed with figitumumab therapy, but it is generally manageable and well tolerated. Because of its extended half-life and absence of dose-limiting toxicity and hypersensitivity, figitumumab compares well to other compounds in its class. Furthermore,

recent data suggest that figitumumab might be active in combination with platinum doublets for the treatment of chemotherapy-naive non-small-cell lung cancer (NSCLC). This article discusses the results to date of the figitumumab development program and the rationale for further testing of this agent as a therapeutic INCB028050 nmr option for the treatment of patients with NSCLC.”
“Recent studies have implicated glial cells in modulation of synaptic transmission, so it is plausible that glial cells may have a functional role in the hyperexcitability characteristic of epilepsy. Indeed, alterations in distinct

astrocyte membrane channels, receptors, and transporters have all been associated with the epileptic state. This review focuses on the potential roles of the glial water channel aquaporin-4 (AQP4) in modulation of brain excitability and in epilepsy. We will review studies of mice lacking AQP4 (Aqp4-/- mice) or a-syntrophin (an AQP4 anchoring protein) and discuss the available human studies demonstrating alterations of AQP4 in human epilepsy tissue specimens. We will conclude with new studies of AQP4 regulation and discuss the potential role of AQP4 in the development of epilepsy (epileptogenesis). While many questions remain unanswered, LY3039478 cell line the available data indicate that AQP4 and its molecular partners may represent important new therapeutic targets. (c) 2012 Wiley Periodicals, Inc.”
“Objective. Altered calcium homeostasis has been linked to increased intima-media thickness (IMT) and plaques. We aimed to investigate whether serum 25-hydroxyvitamin

D (25(OH) D) and serum calcium are associated with IMT and plaques in nonsmoking population. Methods. Ultrasound of the right carotid artery with the measurements of IMT and plaques was performed selleck chemical in 4194 nonsmoking subjects with available measurements of serum 25(OH) D and total calcium. Linear regression was applied to study the linear relationships between variables. Multinomial logistic regression was used to evaluate predictors of increased IMT and total plaque area (TPA), adjusted for age, body mass index, systolic blood pressure, and total cholesterol. Results. There was no significant linear relationship between mean IMT, TPA, and either serum 25(OH) D or total serum calcium. One SD increase in serum 25(OH) D was independently associated with increased odds of being in the highest quartile of IMT in men (OR 1.30, 95% CI 1.12, 1.51). In women, 1 SD increase in serum 25(OH) D was independently associated with increased risk of being in the upper tertile of TPA (OR 1.

\n\nObjective: This study investigated blood and urine heavy metals concentrations, environmental exposure factors, personal behaviors, dietary intakes and the genotypes of related susceptibility genes in patients SB273005 Cytoskeletal Signaling inhibitor with androgenic alopecia (AGA).\n\nDesign: Age, AGA level, residence area, work hours, sleep patterns, cigarette usage, alcohol consumption,

betel nut usage, hair treatments, eating habits, body heavy metals concentrations and rs1998076, rs913063, rs1160312 and rs201571 SNP genotype data were collected from 354 men. Logistic regression analysis was performed to examine whether any of the factors displayed odds ratios (ORs) indicating association with moderate to severe AGA (>= IV). Subsequently, Hosmer-Lemeshow, Nagelkerke R-2 and accuracy click here tests were conducted to help establish an optimal model.\n\nResults: Moderate to severe AGA was associated with the AA genotype of rs1160312 (22.50, 95% CI 3.99-126.83), blood vanadium concentration (0.02, 95% CI 0.01-0.04), and regular consumption of soy bean drinks (0.23, 95% CI 0.06-0.85), after adjustment for age. The results were

corroborated by the Hosmer-Lemeshow test (P = 0.73), Nagelkerke R-2 (0.59), accuracy test (0.816) and area under the curve (AUC; 0.90, 0.847-0.951) analysis.\n\nConclusions: Blood vanadium and frequent soy bean drink consumption may provide protect effects against AGA. Accordingly, blood vanadium concentrations, the AA genotype of rs1160312 and frequent consumption of soy bean drinks are associated with AGA.”
“In this work, an automated screening method for the simultaneous identification and quantitation of 30 representative multiclass drugs (including opiates, cocaine and its main metabolite, cannabinoids, amphetamines and other stimulants in hair samples) has selleck screening library been developed using fast liquid-chromatography time-of-flight mass spectrometry (LC-TOFMS). The identification and quantitation of the drugs were carried out by liquid chromatography using a C(18) column (4.6 x 50 mm) with 1.8 mu m particle size. Accurate mass measurements of ions of interest (typically [M+H](+)) by electrospray time-of-flight

mass spectrometry in the positive ionization mode were used for unambiguous confirmation of the targeted species. Three sample preparation methodologies were evaluated: (a) direct methanolic extraction by sonication, (b) acidic extraction, and (c) alkaline digestion. Direct methanolic extraction showed better recoveries and cleaner extracts. The limits of detection obtained in hair matrix were as low as 5 pg mg(-1) for cocaine and cannabidiol, ranging from 5 to 75 pg mg(-1) for the studied species while the LOQ ranged from 15 to 250 pg mg(-1). The method has been applied to six hair samples from drug consumer volunteers, where the presence of at least one drug was confirmed by accurate mass measurements within 2 ppm (mass error) in most cases.

058, -0.019). This association was also significant even after the adjustment for EI (-0.019; 95% CI: -0.035, -0.003), GNI (-0.035; 95% CI: -0.055, -0.015), and both EI and GNI (-0.017; 95% CI: -0.033, -0.001). The full model showed that around 0.61 of the variation of LE among countries

may be explained by the GPI, EI and GNI per capita. Conclusion: Z-DEVD-FMK The contribution of peace as a global determinant of LE was empirically considerable even after the adjustment for the economic and education levels of countries. This implies that governments should make efforts to settle peace through implementing good governance based on interactions with both public and other countries.”
“As part of the government response to the Deepwater Horizon blowout, a Well Integrity Team evaluated the geologic hazards of shutting in the Macondo Well at the seafloor and determined the conditions under which it could safely be undertaken. Of particular concern was the possibility that, under the anticipated high shut-in pressures, oil could leak out of the well casing below the seafloor. Such a leak could lead to new geologic pathways for hydrocarbon release to the Gulf of Mexico. Evaluating this hazard required analyses of 2D and 3D seismic surveys, seafloor bathymetry, sediment

buy AS1842856 properties, geophysical well logs, and drilling data to assess the geological, hydrological, and geomechanical conditions around the Macondo Well. After the well was successfully capped and shut in on July 15, 2010, a variety of monitoring activities were used to assess subsurface well integrity. These activities included acquisition of wellhead pressure data, marine multichannel seismic profiles, seafloor and water-column sonar surveys, and wellhead visual/acoustic monitoring. HDAC inhibition These data showed that the Macondo Well was not leaking after shut in, and therefore, it could remain safely shut until reservoir pressures

were suppressed (killed) with heavy drilling mud and the well was sealed with cement.”
“Grafted separators, for which poly(ethylene glycol) borate acrylate (PEGBA) was grafted onto polyethylene (PE) separator, were newly prepared by electron beam irradiation. The grafted separators were characterized by FT-IR, energy dispersive X-ray spectrometer (EDS). The morphological changes of the grafted separators were investigated by scanning electron microscopy (SEM). The degree of grafting was increased with irradiation doses. The ionic conductivity of the grafted separator showed the highest value of 6.24 x 10(-4) S cm(-1) at 10 kGy. In addition, its lithium ion transference number and electrochemical stability were enhanced to 0.53 and 4.8 V, respectively owing to anion trapping effect of the grafted unit. The Li ion cells using the grafted separator showed better cycle performances than that using conventional PE separator at various C-rates and high voltage operation conditions.