2 (inter-quartile range, 4.4-16.9) ��g/mg was observed, while the median eGFR was 98 (inter-quartile range, 85-112) mL/min every 1.73 m2. One hundred and eighteen individuals had microalbuminuria (39.7%), including 44 (37.6%) HCV-positive non-diabetic, 58 (49.6%) HCV-positive diabetic, 7 (6.0%) control non-diabetic, and 8 (6.8%) control diabetic inhibitor Cisplatin subjects. Forty five individuals had high ACR (15.8%), including 13 (29.6%) HCV-positive non-diabetic, 25 (56.8%) HCV-positive diabetic, 2 (4.6%) control non-diabetic, and 4 (9.1%) control diabetic subjects. Only 15 (5.3%) had low eGFR, of whom there were 3 (20%) HCV-positive non-diabetic, 7 (46.7%) HCV-positive diabetic, 3 (20%) control non-diabetic, and 2 (13.3%) control diabetic subjects. Of 85 HCV patients who were on treatment, 44 (51.

8%) showed an ETR. Table Table11 demonstrates the study participants�� characteristics by HCV status. HCV-positive individuals had significantly higher ALT levels and higher microalbuminuria. No difference was detected in age, body mass index, urea, creatinine or gender (Table (Table11). Table 1 Study subjects characteristics by HCV status Microalbuminuria Levels of microalbuminuria were significantly higher among HCV-positive individuals than HCV-negative patients (median 9.5 vs 5.9, respectively, Kruskal-Wallis P = 0.017). A significantly higher prevalence of microalbuminuria (defined as albuminuria > upper tertile of the controls) was observed among HCV-positive individuals (53.7%) compared to HCV-negative individuals (31.8%), (��2 = 9.8, P = 0.002). Log microalbuminuria was significantly correlated with grade (r = 0.

13, P = 0.036), borderline correlated with older age (r = 0.11, P = 0.069) and more fibrosis (r = 0.12, P = 0.061), but not significantly associated with viral load (r = -0.03, P = 0.610), or ALT levels (r = -0.03, P = 0.617). There was no significant interaction between HCV status and diabetes such that the odds ratio (OR) of microalbuminuria among diabetics compared to non-diabetics was not significantly higher in HCV-positive individuals compared to HCV-negative controls (P for interaction = 0.720) (Table (Table22). Table 2 Renal insufficiency in diabetics compared with non-diabetics among HCV-positive and HCV-negative controls For sensitivity analysis we restricted the analysis to non-diabetics.

Among non-diabetics, the prevalence of microalbuminuria was significantly higher in HCV-positive GSK-3 individuals (50%) compared to HCV-negative controls (25.8%) (��2 = 5.9, P = 0.015). Restricting the analysis to individuals with no cryoglobulinemia revealed that microalbuminuria was significantly higher in HCV-positive individuals (53.3%) compared to HCV-negative controls (31.8%) (��2 = 9.4, P = 0.002). To adjust for potential confounders and important covariates we employed multivariate regression to test for the significance of HCV as a predictor for microalbuminuria.

For inhibition experiments with various phospholipase inhibitors, after a 24-hour incubation with [3H]AA (as performed and described previously), the cells were washed twice with DMEM/BSA. The washed cells were treated with 1 ��M concentrations of the various inhibitors for Tipifarnib purchase 15 minutes before the addition of peptide #1 (0.1 ��M) and were incubated for an additional 30 minutes, after which the release of [3H]AA was measured by counting the supernatant in a ��-counter. Statistical Analysis. All results are expressed as mean �� S.E.M from at least four experiments. P < 0.05 was considered statistically significant, using analysis of variance (ANOVA, nonparametric) or the paired t test when performed with a specifically matched paired control. Prism GraphPad 4.0 was used for all statistical analyses.

In addition, binding data and monophasic dose-response curves of cellular activation were curve-fitted using Prism (nonlinear curve-fitting). Results Affinities for Human Bn Receptors. In this study, we compared the ability of the naturally occurring Bn-related peptides GRP and NMB and the universal Bn-receptor agonist ligand peptide #1 (Mantey et al., 1997; Pradhan et al., 1998) to interact with and activate the three human Bn-receptor subtypes��hGRP-R, hNMB-R, and hBRS-3 (Jensen et al., 2008)��with that of MK-5046, a recently reported (Guan et al., 2011; Sebhat et al., 2011) BRS-3-receptor selective nonpeptide agonist, and Bantag-1, a BRS-3 selective peptide antagonist (Guan et al., 2010; Feng et al., 2011).

To be certain the results were reflective of the pharmacology of these peptides, two different Bn-receptor�Ccontaining cells were used to assess the results with each human Bn-receptor subtype. In the cells containing hBRS-3, hBRS-3 transfected Balb 3T3 cells (Benya et al., 1995; Mantey et al., 1997) and NCI-N417 lung cancer cells, which contain a native hBRS-3�Creceptor (Ryan et al., 1998b; Uehara et al., 2011), the universal ligand peptide #1 had a high affinity for the hBRS-3-receptor (2.5�C2.7 nM; Fig. 1, A and B; Table 1), similar to what was reported by others (Gonzalez et al., 2008; Sancho et al., 2010; Uehara et al., 2011); this affinity was 2-fold lower than the affinity observed with the antagonist Bantag-1 (1.0�C1.6 nM; Fig. 1; Table 1). However it was 8-fold higher than that for the nonpeptide agonist MK-5046 (17�C18 nM; Fig.

1; Table 1), and >10,000-fold higher than the affinity of GRP and NMB for each hBRS-3 cell line Batimastat (Fig. 1; Table 1). TABLE 1 Binding affinities of different ligands for human BRS-3, GRP, and NMB receptors in various cells Fig. 1. Comparison of the affinities of the peptide agonists GRP, NMB, and peptide #1, nonpeptide agonist MK-5046, and the putative peptide antagonist Bantag-1 for the hBRS-3�Creceptor in (A) hBRS-3 Balb 3T3 cells and (B) NCI-N417 cells. The peptides were … In the two cell lines containing hGRP-R, hGRP-R-transfected Balb 3T3 cells (Benya et al.

Briefly, 10 ��g of tissue or cell lysate or 25 ��l of conditioned medium were boiled for 10 min under reducing conditions and extracts separated on a 10% SDS-polyacrylamide gel, blotted onto nitrocellulose, and stained with 0.5% Ponceau S to assure equal protein loading and transfer. Membranes were blocked with 5% powdered milk and incubated overnight at 4��C with rabbit selleckchem anti-rat MMP-9 (1:1,000, AB 19016; Chemicon, Temecula, CA), followed by the appropriate alkaline phosphatase-conjugated secondary antibody for 2 h (Promega, Madison, WI). Immunodetected proteins were visualized utilizing nitroblue tetrazolium and bromo-chloro-indolyl-phosphate. Standardization was achieved by loading equal amounts of protein (tissue and cell lysates) or using equal cell numbers (cell lysates and conditioned medium).

Cell Isolation and Culture Cell lines. Mouse macrophage cell lines RAW 264.7 (ATCC no. TIB-71) and J774A.1 (ATCC no. TIB-67), human hepatoma cells Huh-7, the rat HSC line CFSC-2G (18) (kindly provided by Dr. M. Rojkind, Washington, DC), and the mouse cholangiocyte cell line 603B (19) were cultured under standard conditions as previously described (32, 33, 35) at 95% air-5% CO2 in a humidified atmosphere. Rat peritoneal macrophages. Rat peritoneal macrophages (pM��) were isolated using a standard protocol (13). Briefly, adult male Sprague-Dawley rat (250 g) were euthanized in a CO2 chamber and injected intraperitoneally through a 20-gauge needle with 50 ml of cold Ca2+, Mg2+-free PBS, followed by aspiration of macrophage-containing medium using the same syringe and centrifugation for 10 min at 400 g.

All procedures were done at 4��C and under sterile conditions. Macrophage yields were 1.4 �� 107 to 2.3 �� 107 cells/rat at a purity of 85% or higher as determined by phagocytosis of 3 ��m latex beads as previously described (35). Apoptosis was induced in vitro in 603B cholangiocytes by a single UV irradiation (200 mJ, Bio-Rad GS Gene Linker UV-chamber; Bio-Rad, Hercules, CA). Apoptotic cells were collected after 3 h, centrifuged at 13,000 g for 5 min, and resuspended in serum-free (1% penicillin/streptomycin; 4 �� 106 cells/ml) before each experiment. Apoptotic cell death was confirmed as previously established (9), assessing cellular criteria (cell rounding, cytoplasmatic membrane blebbing), nuclear morphology (condensed and fragmented nuclei by DAPI staining), and annexin V positivity by fluorescence-activated cell sorting.

It was routinely 80% or higher. For apoptotic cell engulfment experiments, 1�� 104 to 4 �� 104 apoptotic 603B cholangiocytes were added to unstimulated macrophages in serum-free Entinostat medium (freshly isolated rat pM�� or RAW cells, 3 �� 104 cells/well) for 1�C16 h. Determination of ECM-degrading Activities Determination of net interstitial collagenase and gelatinase activities.

stercoralis detection selleck chemicals llc was significantly higher than the sensitivity of the PCR (47.1% versus 17.4%; P < 0.001). The specificity of the Baermann method was 78.4%, and the specificity of PCR was 93.9%. Table 2 Diagnostic accuracy of duplicate Kato�CKatz thick smears, FLOTAC dual technique, and real-time PCR for hookworm and the Baermann method and PCR for S. stercoralis detection as well as prevalence according to three different statistical approaches ... Accuracy estimates of diagnostic methods using a pseudo-gold standard. As shown in Table 2, when applying a combination of the available test results (duplicate Kato�CKatz thick smears, FLOTAC, and PCR) as a diagnostic pseudo-gold standard, the sensitivity for hookworm diagnosis was highest for FLOTAC (83.3%) followed by Kato�CKatz thick smear (75.

0%), and PCR (73.6%). For the diagnosis of S. stercoralis, the Baermann method showed a better sensitivity (83.6%) than PCR (30.9%). Accuracy estimates of diagnostic methods in the absence of a true gold standard using a Bayesian approach. For the comparison of the FLOTAC and Kato�CKatz methods, the two dependence parameters were close to zero, and therefore, we report the following results under the assumption of conditional independence. As shown in Table 2, in the absence of a gold standard, FLOTAC had the highest sensitivity for hookworm detection compared with Kato�CKatz thick smear (96.3% versus 89.6%) and PCR (88.8% versus 83.3%). The sensitivities of Kato�CKatz thick smear (79.2%) and PCR (78.8%) were estimated to be almost equal. The estimated specificity of the PCR was 96.

2% compared with FLOTAC and 92.7% compared with duplicate Kato�CKatz thick smears. For the diagnosis of S. stercoralis, both the Baermann method and PCR showed low sensitivity (28.3% and 11.6%, respectively). The specificity of PCR was higher than the specificity of the Baermann method (90.6% versus 75.2%). Discussion The upscale of control interventions against neglected tropical diseases over the next years in accordance with the WHO goals set for the year 2020 will likely reduce the prevalence and intensities of soil-transmitted helminth infections in endemic countries. For the decisions of where to implement, when to stop control interventions, and how to implement adequate surveillance to avoid the recrudescence of soil-transmitted helminthiases, sensitive diagnostic methods are needed.

52 For the first time GSK-3 to our knowledge, we compared the diagnostic accuracy of the FLOTAC with a previously described real-time PCR assay for hookworm diagnosis, applying three different statistical approaches, and we found that FLOTAC was slightly more sensitive than PCR. When directly comparing each of the techniques with the Kato�CKatz method, only FLOTAC and not PCR had a significantly higher sensitivity. The sensitivity of the PCR for S. stercoralis diagnosis was significantly lower than the sensitivity of the Baermann method.

2]. As the vaccine efficacy is only 85% for first dose of measles at nine completed months, there selleck chem is progressive accumulation of a small number of susceptible children in the community over the years. Such accumulations are typically caused by the combination of the measles vaccine efficacy that does not reach 100% and children left un-immunized each year. As the coverage increases, inter-epidemic interval increases as well as a shift towards older age groups may be observed as in Thailand and Sri Lanka.[18] Although awareness and practice regarding measles were satisfactory among healthcare providers in both settings yet in the Shahpur block workers, there were observable gaps not only in the knowledge part but also in the scientific usage of the vaccine carriers, while conducting the immunization sessions in the fields which might lead towards the mechanism of cold chain failure.

Yeung et al. recorded the findings of the cold chain failure despite 95% vaccination as one of the factors for the measles outbreak.[19] With respect to beneficiaries related issues, results in two areas suggest difference with respect to knowledge regarding cause of measles, treatment and follow-up practices. Measles is locally known as Dharrssali. Illiteracy and knowledge are complimentary. Added with beliefs and barriers, many statistically significant factors like geographically difficult hilly areas, poor socio-economic status, poverty with over-crowding in the muddy thatched houses and illiterate mothers were found to be more inclined towards traditional unscientific lines in terms of cause and effect.

Earlier studies had also shown that cases hypothesize the genesis of measles as curse of goddess.[20�C23] Despite high immunization coverage in the areas, majorities of respondents opted for traditional faith healing treatment and with diet rich in Seul�Ca medicinal plant employed to facilitate eruption of measles during illness. For the vaccination of the children, they had to travel a distance of6�C10 kms on hilly terrain on foot consuming 3/4th hour to 2 and ? hour for nearest available health care facility [Table 4]. The attack rates even among highly vaccinated children aged 11�C17 years in Jathrear and 6�C15 years in Kutharna villages were higher. This suggests waning of immunity with age (secondary vaccine failure), which can be due to the use of poorly stored vaccine.

However, effect of exposure to ultra Batimastat violet radiation to wane immunity as observed by Mary cannot be ruled out.[24] There was no measles cases in the age group of 0�C5 years, and it is possible that there is higher risk of the occurrence of secondary vaccine failure that reduce the immunity to measles over time than the primary vaccine failure that did not develop the immunity. Also, there is the possibility of the failure of the cold chain maintenance in the area with the lower vaccine efficacy at the time when the older children aged 11�C16 years in Jathrear had their measles vaccine.

5.3. GATKAs described in the variant calling section, the Genome Analysis ToolKit (GATK), which provides selleck inhibitor a collection of data analysis tools, can also allow indel calling based on the MapReduce programming approach [22]. Details of GATK in comparison to other indel calling methods including Dindel (VarScan, SAMtools mpileup) are evaluated in Neuman et al. [38].6. Filtering and AnnotationAfter alignment and variant calling, a list of thousands of potential differences between the genome under study and the reference genome is generated. The next step is to determine which of these variants are likely to contribute to the pathological process under study. The third step involves a combination of both filtering (removing variants that fit specific genetic models or are not present in normal tissue) as well as annotation (looking up information about variants and identifying ones that fit the biological process).

Filtering can be done with a genetic pedigree or with cancer and normal samples from the same individual. In the instance of cancer, a common method is removing variants that are present in both the cancer sample and the normal sample, leaving only somatic variants, which have mutated from the germline sequence. In the instance of a pedigree, filtering can be done based on the different inheritance patterns. For example, if the inheritance pattern is autosomal recessive, the variants that are heterozygous in the parents and homozygous in the child can be chosen. Similar methods can be done with larger pedigrees based on the inheritance pattern.

In addition to filtering, further selection of causal variants can be based on existing annotation or predicted functional effect. Many tools exist to examine relevant variants by referencing previously known information about their biological functions and inferring potential effects based on their genomic context. In addition, many tools have been developed to identify genetic variants that cause disease pathogenesis or phenotypic variance [39]. Rare nonsynonymous SNPs are SNPs that cause amino acid substitution (AAS) in the coding region, which potentially affect the function of the protein coded and could contribute to disease.The advance of exome and genomic sequencing is yielding an extensive number of human genetic variants, and a number of disease-associated SNVs can be identified following alignment and variant calling.

AV-951 Unlike nonsense and frameshift mutations, which often result in a loss of protein function, pinpointing disease-causal variants among numerous SNVs has become one of the major challenges due to the lack of genetic information. For instance, ~1,300 loci are shown to be associated with ~200 diseases by GWASs but only a few of these loci have been identified as disease-causing variants [40].

An opposite trend was observed in the olive paste at 200DAF in which it was found that (E)-2-hexenal content significantly increased www.selleckchem.com/products/INCB18424.html after 30min of malaxation time.In 230DAF fruits, no significant variations in (E)-2-hexenal concentration for both areas of cultivation were observed. This trend may be due to the different degree of inactivation of enzymes. In fact, LOX enzyme can be influenced by the different phenolic levels monitored in the fruit at the various stages of ripening. The inactivating role of phenolic compounds on enzyme activity is well established [36]. In the olive oils obtained from 170DAF fruits in each area, phenolic compounds were significantly greater in 170DAF with respect to 200 and 230DAF fruits (Table 2).3.4.

Other ParametersPhenols and lipids contribute to determine the nutritional qualities of vegetable oils, but the fatty acid composition is also very important. In fact, monounsaturated fatty acids, such as oleic acid, exert beneficial effects in cardiovascular disease prevention [37]. Consequently, the increase of oleic acid has become one of the major goals to improve vegetable oil quality. However, it is the presence of minor components, in particular phenols, contributing to oil’s high oxidative stability, to colour and flavour, that makes olive oil unique among other oils.The pattern of the parameters investigated (Table 2) showed that the ripeness stage of ��Coratina�� olives that yields the best quality of oil corresponds to the 200DAF fruits. In fact, the oils produced by olives harvested at this time frame (200DAF fruits) are characterized by high nutritional properties (i.

e., oleic acid and total phenol content).4. ConclusionsThe results show that the LOX transcript accumulation increased during olive fruit ripening on the ��Coratina�� genotype cultivated in two different farms. This trend suggests that it is connected with ripening and senescence processes. Dacomitinib In Mirto-Crosia farm, the LOX transcripts in olive fruits were significantly more abundant than in the Rende farm. The difference of LOX transcript accumulation between the Mirto-Crosia and Rende farms may be due to environmental differences (i.e., temperature, soil fertility, humidity, etc.). Olive paste and olive oil samples, obtained by fruits collected in both farms, showed different volatile contents. The volatile molecules are produced via the LOX pathway during the mechanical oil extraction process and represent the main classes of compounds responsible for ��fruity��, ��cut-grass��, and ��floral�� flavours. Although the response of the volatile compound concentrations depended mainly on the growing season, there were some consistent trends which could be attributed to environmental differences.

The method uses an intermediate model that acts as bridge between Pacritinib the annotated UML models and the analysis-oriented models. In particular, discrete time Markov process models can be derived for the computation of the service reliability.Finally, the work [28] proposes a model-to-model transformation technique to support the availability evaluation of railway control systems. The availability model is a repairable fault tree that is automatically generated from the UML models (use case, component, and state machine diagrams), properly annotated with MARTE and DAM extensions.8. Conclusion A standard specification framework is yet needed for dependability assessment of UML-based specifications. DAM is a step toward this goal, as it is a comprehensive approach attempting to unify a great number of efforts carried out by the researchers in the last decade.

Software quality includes a number of very different NFPs (e.g., security, performance, and dependability), which are often in conflict with each other [29]. In this context, the MARTE-DAM profile is a promising common framework for the specification of different NFPs in UML-based design. We envisage that a future research goal is to devise model transformation techniques that support a comprehensive analysis in presence of conflicting NFPs (e.g., performability, vulnerability, and survivability issues), in order to provide trade-off solutions to the software engineer. Acknowledgments This paper has been partially supported by Fundaci��n Arag��n I+D, by Spanish Projects TIN2011-24932 and DPI2010-20413 and by a Discovery grant from the Natural Sciences and Engineering Research Council of Canada (NSERC).

AppendixA. MARTE and DAM Profiles A.1. MARTE The ��UML Profile for Modeling and Analysis of Real-Time and Embedded systems�� [12] is an OMG standard profile that extends UML in a lightweight fashion (i.e. via the standard UML extension mechanism including stereotypes, tagged values, and constraints) and enables the specification of both quantitative and qualitative nonfunctional properties (NFP) in the form of annotations attached to UML model elements.The Value Specification Language (VSL), which is a part of MARTE, provides the ability to express NFP types (defined in the so called MARTE library), values of NFP types, as well as variables, constants, and expressions.

All of these are used by the modeler to assign values to tags, according to the VSL syntax (the annotations in Figure 2 show several examples). Tags of NFP types are characterized by several qualifiers: source defines the origin of the specification (such as required, assumed, predicted, and measured); statQ defines the type of a statistical measure (such as a maximum, Cilengitide minimum, or mean); unit indicates the measurement unit for a given NFP.

Moreover, one of the selleckbio challenges in the treatment with sorafenib is the difficulties in assessing tumor response by traditional response criteria. The pivotal trial by Llovet et al. reported a very modest response rate of only 2% [4]. The lack of a correlation between objectively observed response and clinical benefit complicates treatment evaluation and clinical decision making [7]. A clinical improvement in patients’ symptoms may not be expected, as no significant difference between time to symptom worsening has been observed in sorafenib-treated patients compared to placebo [4]. Recent studies indicate that an early decline in serum ��-fetoprotein (��FP) may be a predictive marker for treatment response to targeted therapies in advanced HCC [7, 8].

In the current study we evaluate the efficacy and tolerability of sorafenib in an unselected patient population as they present in every day clinical practice. Furthermore we explore the role of ��FP in treatment evaluation and its correlation to survival outcome.2. Materials and Methods2.1. PatientsAccess to sorafenib was made available in August 2007 through a program under the Danish National Board of Health. All patients considered for sorafenib were reviewed by a panel of experts appointed by the National Board of Health and referred to one of two centrs designated to treat HCC patients with sorafenib in Denmark.

A common set of criteria for the selection of patients were used by the two centres: advanced hepatocellular carcinoma diagnosed according to the criteria of EASL [9], not amendable for locoregional treatment (including transcatheter arterial chemoembolization, radio frequency ablation (RFA), and surgery), ECOG PS 0�C2, CP A or B, and no Drug_discovery substantial co-morbidity (uncontrolled cardio- or cerebrovascular disease, recent bleeding episodes, or active ulcer disease).All patients had a dynamic three-phase CT scan performed at baseline as well as an electrocardiogram, blood pressure measurement, blood samples including haematological values, liver biochemistry, and serum ��FP.2.2. TreatmentSorafenib was administered at a dose of 800mg daily. However, weak or elderly patients started at a reduced dose of 400mg daily, with the possibility of dose escalation. Dose reduction and treatment delay were performed according to the recommendations in the summary of product characteristics [2].Treatment was continued until radiological or clinical progression, unacceptable toxicity, death, or patient refusal. Patients were seen every 4 weeks for toxicity management and clinical assessment. Response evaluation was performed every 12 weeks and included a CT-scan, liver biochemistry, and serum ��FP. 2.3.

For determination of ROS scavenging capacity, the H2O2 content of lime plants under EMF stress was estimated. H2O2 content was always significantly lower under EMF stress throughout the experiments until performed here (Figure 6). This result is in contrast to the obtained result on MDA content (Figure 5). Lower content of H2O2 might be a result of the significantly higher induction of the activities of antioxidant enzymes in the lime plants under EMF stress. Several authors have described the overproduction of toxic oxygen forms with aging in plants [37, 38]. It is known that ROS can cause peroxidation of membrane fatty acids [39]. In turn, these oxidized fatty acids may give rise to the propagation of peroxidation resulting in membrane damage.

In a recent report, Lacan and Baccou [40, 41], besides supporting that ROS are involved in the ripening and senescence events in nonnetted muskmelon fruit, also showed that the observed delay in senescence in the long-storage life variety Clipper is closely linked to the very low free radical induced membrane lipid peroxidation in comparison with that of the short-storage life variety Jerac. These data explain the EMF-improved ability to scavenge free radicals, leading to a delay in senescence and alterations in membrane integrity, as demonstrated by the growth and survival responses. Furthermore, these results suggest that, under specific conditions, it takes the combined action of more than one antioxidant to provide an increased resistance to oxidative stress and lengthened survival in plants. This is in agreement with some previous reports [42, 43].

Figure 6Effect of EMF on H2O2 content in healthy and infected plants of Citrus aurantifolia. Vertical bars indicate mean �� SE of three replicates. Different letters indicate significant differences (P < 0.05).4. ConclusionsThe obtained result show that 10KHz EMF field can simulate the growth rate of healthy and infected lime plants. Moreover, it seems that EMF field could reduce the intensity of Witches' broom disease in infected trees and this effect could probably be due to the reduction of phytoplasma in plant tissues. This study provides an initial understanding of the response of infected lime plants to EMF stress, which is important for future studies aimed at developing strategies for struggle against phytoplasma and Witches' broom disease in lime plants.AcknowledgmentsThe financial support of this research was provided partly by College of Science, University of Tehran and partly by Engineering Research Institute.
In higher plants, salinity GSK-3 can provoke alterations in the metabolism of proteins and nucleic acids, photosynthesis and respiration [1�C3].