The Sig1R agonist 4-IBP and related antagonist BD1047 Ruboxistaurin chemical structure were also inhibitory. The Sig1R agonist SKF10047 had no effect. Sustained calcium entry evoked by VEGF or hydrogen peroxide was also inhibited by BD1063, BD1047 or 4-IBP, but not SKF10047. 4-IBP, BD1047 and BD1063 inhibited TRPC5 or TRPM3, but not TRPM2. Inhibitory effects of BD1047 were rapid in onset and readily reversed on washout. SKF10047 inhibited TRPC5 but not TRPM3 or TRPM2. Depletion

of Sig1R did not prevent the inhibitory actions of BD1063 or BD1047 and Sig1R did not co-localize with TRPC5 or TRPM3. Conclusions and Implications The data suggest that two types of Sig1R ligand (BD1047/BD1063 and 4-IBP) are inhibitors of receptor- or chemically activated calcium entry channels, acting relatively directly and independently of the Sig1R. Chemical foundations for TRP channel inhibitors are suggested.”
“Background: Data regarding the patterns and the mechanisms of deregulation of the insulin growth factor (IGF) pathway in adult and paediatric gastrointestinal NSC23766 purchase stromal tumours (GISTs) are

limited.\n\nMethods: We investigated the expression profiling of the genes encoding the main components of the IGF signalling pathway in 131 GISTs (106 adults, 21 paediatric and four young adults) and 25 other soft-tissue sarcomas (STS) using an Affymetrix U133A platform. IGF2 was investigated for loss of imprinting (LOI) whereas IGF1R was analysed for copy number aberration and mutation.\n\nResults: IGF2 was the most highly overexpressed gene of the IGF pathway in GIST. IGF2 expression was also significantly higher than in other STS. IGF2 expression was correlated to the age onset and mutational status of GIST. Indeed, IGF2 expression was significantly higher in the ‘adult’ group than in the ‘paediatric’ and ‘young

adult’ groups. Among adult GIST, IGF2 expression was higher in tumours lacking Homo sapiens v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (KIT) or alpha-type platelet-derived growth factor receptor (PDGFRA) mutations in comparison with mutated cases. A trend for a higher expression of IGF2 in resistant GIST SCH727965 concentration in comparison to responsive GIST was also found. Overexpression of IGF2 was not related to LOI. Conversely, the expression of the IGF1R gene was significantly higher in the paediatric group than in the adult group. No copy number gains or mutations of IGF1R were observed.\n\nConclusion: The IGF pathway is deregulated in GIST with distinct patterns according to age onset and mutational status. The IGF pathway may represent a therapeutic target in patients with primary or secondary resistance to imatinib. (C) 2012 Elsevier Ltd. All rights reserved.”
“A novel type of anion channel activated by extracellular acidification, called acid-sensitive outwardly rectifying (ASOR) anion channel, was shown to be involved in acidotoxic necrotic death in human epithelial cells.

“
“We report on non-equilibrium molecular dynamics calculations of chiral single-wall carbon nanotubes using the framework of Objective Structures. This enables us to adapt molecular dynamics to the symmetry of chiral nanotubes and efficiently simulate these systems with small unit

cells. We outline the method and the adaptation of a conventional thermostat and barostat to this setting. We then apply the method in order to examine the behavior of nanotubes with various chiralities subject to a constant extensional strain rate. We examine the effects of temperature, strain rate, and pre-compression/pre-tension. We find a range of failure mechanisms, find more including the formation of Stone-Wales defects, the opening of voids, and the motion of atoms out of the cross-section. (C) 2011 American Institute of Physics. [doi:10.1063/1.3596827]“
“Title. Complex decisions: theorizing women’s infant feeding decisions

in the first 6 weeks after birth.\n\nAim. This paper is a report of a grounded theory study of woman’s infant feeding experiences and decisions in the first 6 weeks after birth.\n\nBackground. Breastfeeding is considered the optimum method of infant feeding. Studies have identified numerous factors associated with infant feeding decisions. What remains unexplored are the mechanisms by which socio-demographic, biomedical and psychosocial factors influence IPI-145 clinical trial infant feeding decisions. Research highlights the need for further investigation of the experiences and decision-making processes of both breastfeeding and formula-feeding women.\n\nMethod. A constructionist grounded theory approach to data collection and analysis was used. Data for this study were collected and analysed between 2003 and 2004. Further data,

collected in a previous study in 2000, were theoretically sampled and analysed in 2005. In-depth interviews with 37 women from various socio-demographic areas in New South Wales, Australia provided data.\n\nFindings. The core category was “deconstructing best’. “Deconstructing best’ was the infant feeding decision-making process in the first 6 weeks after birth. The process of “deconstructing best’ involved seven phases: planning, expecting, realizing, questioning, getting on with it, defending and qualifying. ALK inhibition Four main categories – ‘it’s really best to breastfeed’, ‘it’s the unknown’, ‘it’s not the only thing going on’, and ‘everybody’s best is different’ – comprised the context within which deconstructing best occurred.\n\nConclusion. Woman’s infant feeding decisions cannot be viewed in isolation from other post-natal experiences and needs. Infant feeding decisions will only be understood and appropriately supported when they are seen in relation to the circumstances of a woman’s life, her immediate sociocultural context and individual experience.”
“Sagebrush (Artemisia spp.

These results suggested that phyloproteomics might be a useful tool for the phylogenetic analysis of tick species in which sequence data is a limiting factor and demonstrate the possibilities of proteomics studies for the characterization of relevant

tick vector species and provide new relevant information to understand the physiology, development and evolution of these tick species. Biological significance This is the first report on the proteome of the most important Amblyomma tick species for their relevance as vectors of zoonotic pathogens worldwide. Nevertheless, very little information learn more is available on the genome, transcriptome and proteome of these vector ectoparasites. The results reported herein provide new relevant

information to understand the physiology, development and evolution of these tick species. Phyloproteomics using de novo protein sequencing was assayed as a new approach for the phylogenetic analysis of tick species in which sequence data is a limiting factor. This article is part of a Special Issue entitled: Proteomics of non-model organisms. (C) 2014 Elsevier B.V. All rights reserved.”
“OBJECTIVE. The purpose of this study was to evaluate interobserver agreement and whether or not a new MRI grading system correlates with symptoms and neurologic signs for assessing spinal canal compression.\n\nMATERIALS AND METHODS. One hundred patients (52 men and 48 women; mean age, 50 years) underwent MRI of the cervical www.selleckchem.com/products/z-ietd-fmk.html spine at our institution and were evaluated by two musculoskeletal radiologists. The presence and grade of cervical canal stenosis at the maximal narrowing point was assessed according to the new grading system suggested by Kang et al. (Kang system). The results correlated with the clinical manifestations and neurologic examination. Statistical analysis was performed using kappa statistics, categoric regression analysis, and nonparametric correlation analysis (Spearman correlation).\n\nRESULTS. Interobserver agreement in the grading of spinal stenosis between the two readers was almost perfect (kappa = 0.925). Most of the patients with grade 0 cervical

canal stenosis showed no neurologic manifestation, and patients with grades 2 and 3 EPZ-6438 solubility dmso cervical canal stenosis had positive neurologic manifestations. The correlation coefficient (R) of reader 1 between MRI grade (0, 1, 2, and 3) and neurologic manifestations (positive or negative) was 0.846. The R of reader 2 was 0.808. In the younger age group (< 50 years old), the R of reader 1 was 0.834 and the R of reader 2 was 0.745. In the older age group (>= 50 years old), the R of reader 1 was 0.839 and the R of reader 2 was 0.839.\n\nCONCLUSION. The interobserver agreement of the Kang system was almost perfect and was higher than in the study by Kang et al. Grade 0 cervical canal stenosis represents negative neurologic manifestations and grades 2 and 3 cervical canal stenosis represent positive neurologic manifestations.

\n\nTwo hundred and sixty-two patients met inclusion criteria. VUR was detected in 21.3 %, urologic abnormality including VUR in 27.4 %. Degree of bladder distension, department of referring physician, study indication, positive documented urine culture, and history of recurrent UTI or UTI and other

check details abnormality were all not associated with increased likelihood of VUR or any urologic abnormality on VCUG.\n\nVUR and VCUG abnormality are no more likely when performed after recurrent UTI or for UTI plus other abnormality. This reasons against postponing VCUG until after UTI recurrence, as positive findings are no more likely in this setting.”
“An atmospheric-pressure (AP) argon plasma discharge generated in a single-electrode configuration with the power supply operating at a frequency of 45 kHz is employed to perform some applications, such as the treatment of the interior surface of a medical infusion tube, hydrophilic modification of insulator surface, hardening of metal surface, and acidification treatment of water. It is shown that the active gases (O-2 and N-2) mixed in a carrier gas (Ar) play a significant role in carrying out such applications. A preliminary study on its applications is presented to demonstrate the promising potential selleck compound of AP single-electrode-configuration plasma jet.”
“G-quadruplex-forming oligonucleotides

containing modified nucleotide chemistries have demonstrated promising pharmaceutical potential. In this work, we systematically investigate the effects of sugar-modified guanosines on the structure and stability of a (4+0) parallel and a (3+1) hybrid G-quadruplex using over 60 modified sequences containing a single-position substitution of 2′-O-4′-C-methylene-guanosine ((LNA)G), 2′-deoxy-2′-fluoro-riboguanosine ((F)G) or 2′-deoxy-2′-fluoro-arabinoguanosine ((FANA)G). Our results are summarized in two parts: (I) Generally, (LNA)G substitutions into selleck kinase inhibitor ‘anti’ position guanines within a guanine-tetrad

lead to a more stable G-quadruplex, while substitutions into ‘syn’ positions disrupt the native G-quadruplex conformation. However, some interesting exceptions to this trend are observed. We discover that a (LNA)G modification upstream of a short propeller loop hinders G-quadruplex formation. (II) A single substitution of either (F)G or (FANA)G into a ‘syn’ position is powerful enough to perturb the (3+1) G-quadruplex. Substitution of either (F)G or (FANA)G into any ‘anti’ position is well tolerated in the two G-quadruplex scaffolds. (FANA)G substitutions to ‘anti’ positions are better tolerated than their (F)G counterparts. In both scaffolds, (FANA)G substitutions to the central tetrad layer are observed to be the most stabilizing. The observations reported herein on the effects of (LNA)G, (F)G and (FANA)G modifications on G-quadruplex structure and stability will enable the future design of pharmaceutically relevant oligonucleotides.

Results Fifty-three per cent of the cohort was male. Eighty-nine per cent were outpatients. Bowel cleansing was reported as satisfactory/good in 87% and poor in 13%. A <8-h preparation to procedure time was associated with a higher rate of

satisfactory/good cleansing than a >8-h interval (odds ratio (OR) 1.3, P = 0.04). In a multivariate analysis, female gender (OR 1.4, P = 0.02), outpatient status (OR 3.1 P = 0.001) and indication for procedure (P < 0.01) were significant predictors of adequate bowel preparation. Adequate bowel preparation was associated with a significant 5-Fluoracil chemical structure increase in caecal intubation rates (OR 5.3, P = 0.001). Conclusions A shorter (<8h) interval between end of

bowel preparation and start of colonoscopy yielded better bowel cleansing than a longer (>8h) interval. Adequate bowel preparation led to improved caecal intubation rates.”
“In modern drug discovery, numerous assay formats are available to screen and quantitate receptor-ligand interactions. Radioactive assays are “gold standard” because they are fast, easy, and reproducible; however, they are hazardous, produce radioactive waste, require special lab conditions, and are expensive on a large scale. Thus, it provides a lot of importance to the “mix & measure” assays that have an optical readout. Fluorescence techniques are likely to be among the most important detection approaches used for high throughput screening due to their high sensitivity and amenability to automation. The aim of the present study was to determine the functional antagonistic click here affinities of standard muscarinic antagonists in CHO cells over expressing m1, m3, and m5 receptors and to compare

them with the respective binding affinities. This study was further extended to elucidate that Ca+2 measurement assays can serve as a functional screening tool for GPCRs. For this purpose, standard muscarinic receptor antagonists, namely, tolterodine, oxybutynin, and atropine were used. We determined and compard the IC50 values of these https://www.selleckchem.com/products/VX-680(MK-0457).html three standard inhibitors in fura 2 AM loaded m1, m3, and m5 overexpressing CHO cells and in radioligand binding assay. Both the assays exhibited comparable rank order potencies of the standard inhibitors. This study suggests that Ca+2 mobilization assays can be an alternate to radioligand binding assays.”
“This study aimed to assess the efficacy and safety of combination treatment with lenalidomide and cetuximab in KRAS-mutant metastatic colorectal cancer patients. This was a phase II multicenter, open-label trial comprising a safety lead-in phase (phase IIa) to determine the maximum tolerated dose, and a randomized proof of concept phase (phase IIb) to determine the response rate of lenalidomide plus cetuximab combination therapy.

G + C mol%

of strain JC230(T) was 54.1%. Distinct morphological, physiological and genotypic differences from the previously described taxa support the classification of strain JC230(T) as a representative of a new species in the genus Spirochaeta, for which the name Spirochaeta lutea sp. nov. is proposed. The type strain is JC230(T) (=KCTC 15387(T) = DSM 29074(T)). Selonsertib mouse (C) 2014 Elsevier GmbH. All rights reserved.”
“TRAIL (TNF-related apoptosis-inducing ligand) is a promising anticancer agent that can be potentially used as an alternative or complementary therapy because of its specific antitumor activity. However, TRAIL can also stimulate the proliferation of cancer cells through the activation of NF-kappa B, but the exact mechanism is still poorly understood. In this study, we show that chronic exposure to subtoxic concentrations of TRAIL results in acquired resistance. This resistance is associated with the increase in miR-21, miR-30c, and miR-100 expression, which target tumorsuppressor genes fundamental in the response to TRAIL. Importantly, down-regulation of caspase-8 by miR-21 blocks receptor interacting GSK3235025 inhibitor protein-1 cleavage and induces the activation of NF-kappa

B, which regulates these miRNAs. Thus, TRAIL activates a positive feedback loop that sustains the acquired resistance and causes an aggressive phenotype. Finally, we prove that combinatory treatment of NF-kappa B inhibitors and TRAIL is able to revert resistance and reduce tumor growth, with important consequences for the clinical practice.”
“Ishida K, Matsumoto T, Taguchi K, Kamata K, Kobayashi T. Mechanisms underlying altered extracellular nucleotide-induced contractions in mesenteric arteries from rats in later-stage type 2 diabetes: effect of ANG II type 1 receptor antagonism. Am J Physiol Heart Circ Physiol 301: H1850-H1861, 2011. First published August 19, 2011; doi:10.1152/ajpheart.00502.2011.-Little

is known about the vascular contractile responsiveness to, and signaling pathways for, AC220 extracellular nucleotides in the chronic stage of type 2 diabetes or whether the ANG II type 1 receptor blocker losartan might alter such responses. We hypothesized that nucleotide-induced arterial contractions are augmented in diabetic Goto-Kakizaki (GK) rats and that treatment with losartan would normalize the contractions. Here, we investigated the vasoconstrictor effects of ATP/UTP in superior mesenteric arteries isolated from GK rats (37-42 wk old) that had or had not received 2 wk of losartan (25 mg.kg(-1).day(-1)). In arteries from GK rats (vs.

Patients with DO (54.1%) were older (61.8 vs. 50.8 years) and had smaller maximum voided volumes per void (377 mL vs. 476 mL), average 24-h urine output (1,975 mL vs. 2,320 mL), and significantly more incontinent episodes. On UDS, patients with DO were more likely to have abnormal sensation, with strong desire and urgency occurring at significantly lower bladder volumes.\n\nDespite similar symptomatology, there are objective differences between OAB patients with and without DO.”
“In many paediatric cardiac units chest radiographs are performed routinely before discharge after cardiac surgery. These radiographs contribute to radiation p53 inhibitor exposure. To evaluate the

diagnostic impact of routine chest X-rays before discharge in children undergoing open heart surgery and to analyze certain risk factors predicting pathologic findings. This was a prospective (6 months) single-centre observational clinical study. One hundred twenty-eight consecutive children undergoing heart surgery underwent biplane chest X-ray at a mean of 13 days after surgery. Pathologic findings

on chest X-rays were defined as infiltrate, atelectasis, pleural effusion, pneumothorax, or signs of fluid overload. One hundred nine asymptomatic children were included Smoothened Agonist research buy in the final analysis. Risk factors, such as age, corrective versus palliative surgery, reoperation, sternotomy versus lateral thoracotomy, and relevant pulmonary events during postoperative paediatric intensive care unit (PICU) stay, were analysed. In only 5.5 % (6 of 109) of these asymptomatic patients were pathologic findings on routine chest X-ray before discharge found. In only three of these cases (50 %), subsequent noninvasive medical intervention (increasing diuretics) was needed. All six patients had relevant pulmonary events during their PICU stay. Risk factor analysis showed only pulmonary complications during PICU stay to be significantly associated (p = 0.005) with

pathologic X-ray findings. Routine chest radiographs before buy BVD-523 discharge after cardiac surgery can be omitted in asymptomatic children with an uneventful and straightforward perioperative course. Chest radiographs before discharge are warrantable if pulmonary complications did occur during their PICU stay, as this is a risk factor for pathologic findings in chest X-rays before discharge.”
“Background Preterm prelabour rupture of membranes (PPROM) is associated with increased risk of maternal and neonatal morbidity and mortality. Women with PPROM have been predominantly managed in hospital. It is possible that selected women could be managed at home after a period of observation. The safety, cost and women’s views about home management have not been established. Objectives To assess the safety, cost and women’s views about planned home versus hospital care for women with PPROM.

Conclusion: This study indicates that sex hormones induce MetS in a relatively low proportion of healthy

GD individuals and especially during the first year of hormonal treatment. Most importantly, concomitant psychiatric problems are associated with considerably greater MetS prevalence in hormone treated GD individuals. (C) 2015 Elsevier Inc. All rights reserved.”
“Sample processing is often ignored during analytical method development and validation, but accurate results for real samples depend on all aspects of the analytical process. Also, validation is often conducted using only spiked samples, but extraction yields may be lower in incurred samples. In this study, different BMS-777607 ic50 variables in extraction for incurred pesticides and environmental contaminants in fish were investigated. Among 207 analytes screened using low-pressure gas chromatography-tandem mass spectrometry, consisting Panobinostat of 150 pesticides,

15 polycyclic aromatic hydrocarbons (PAHs), 14 polychlorinated biphenyls (PCBs), 6 polybrominated diphenyl ethers (PBDEs), and 22 other flame retardants (FRs), 35 (16 pesticides, 9 PCBs, 5 PBDEs, and 5 PAHs) were identified for quantification in samples of salmon, croaker, and NIST Standard Reference Material 1947 (Lake Michigan Fish Tissue). Extraction efficiencies using different extraction devices (blending, vortexing, and vibrating) versus time, sample size, and sample/solvent ratio were

determined. In comparison to blending results, use of a pulsed-vortexer for 1 min with 1/1 (g/mL) sample/acetonitrile ratio was generally sufficient CYT387 JAK/STAT inhibitor to extract the incurred contaminants in the homogenized fish tissues. Conversely, extraction with a prototype vibration shaker often took 60 min to achieve 100% extraction efficiency. A main conclusion from this study is that accurate results for real samples can be obtained using batch extraction with a pulsed-vortexer in a simple and efficient method that achieves high sample throughput.”
“Seven new iodo-bridged binuclear platinum(II) complexes [Pt(L)I(2)](2) (1-7) (L = n-butylamine, isopropylamine, m-toluidine, p-toluidine, diethylamine, N-methylaniline and aniline) have been synthesized and characterized by elemental analysis, conductivity, thermal analysis, IR. (1)H NMR and mass spectra techniques. The cytotoxicity was tested by MTT assay. The results indicate that they have selectivity against tested carcinoma cell lines. For example, the complexes (3 and 7) have better cytotoxicity than cisplatin against Bel-7402 cell line, the complexes (2-5 and 7) have better cytotoxicity than cisplatin against Hela cell line. The results suggest that the species of amine has important effect on cytotoxicity, when L = m-toluidine, p-toluidine and aniline, the complexes have better cytotoxicity against tested carcinoma cell lines.

Breathing H2S also induces a rapidly reversible reduction of metabolic rate at either body temperature.”
“P>This report describes the isolation of rodent multipotent adult progenitor cells (MAPCs) and proliferation of these cells in both standard medium and medium without exogenous serum or growth factors conditioned by the rat cell line B104. MAPCs have exacting requirements AZD2014 purchase for their proliferation in vitro but once established proliferate rapidly at low seeding density, requiring almost daily passage and media exchange. Previously published methods for growth of MAPCs in vitro all used media supplemented with serum and growth factors, which

adds considerable expense.”
“The adult mammalian heart has an extremely limited capacity for regeneration. As a consequence, ischemic heart disease remains the leading cause of death in the developed world, and the heart continues to be a major focal point for regenerative medicine. Understanding innate mechanisms of heart regeneration is important and may provide a blueprint for clinical translation. For example, urodele amphibians and teleost fish can mount an endogenous regenerative response following multiple forms of cardiac injury, and this

regenerative response appears to be mediated through proliferation of pre-existing cardiomyocytes. How and why mammals have lost the capacity for heart regeneration since the divergence from teleost fish more than 450 million years ago has been a major unresolved question in the field. Recent studies in mice indicate www.selleckchem.com/products/epacadostat-incb024360.html that the mammalian heart possesses significant regenerative potential during embryonic and neonatal life, but this regenerative capacity is lost rapidly after birth. This review focuses on mechanisms of heart regeneration in neonatal mice, with a particular emphasis on similarities and differences with the zebrafish model.

Recent advances in our understanding of the molecular mechanisms of postnatal heart maturation and regenerative arrest are also highlighted. The possibility of recapitulating ontogenetically and phylogenetically ancient mechanisms of cardiac regeneration in the adult human heart represents an exciting new frontier in cardiology. (Trends Cardiovasc Med 2012;22:128-133) (c) 2012 Elsevier Inc. All rights reserved.”
“The 26S proteasome operates at the executive end of the ubiquitin-proteasome pathway. Here, JQ1 manufacturer we present a cryo-EM structure of the Saccharomyces cerevisiae 26S proteasome at a resolution of 7.4 angstrom or 6.7 angstrom ( Fourier-Shell Correlation of 0.5 or 0.3, respectively). We used this map in conjunction with molecular dynamics-based flexible fitting to build a near-atomic resolution model of the holocomplex. The quality of the map allowed us to assign alpha-helices, the predominant secondary structure element of the regulatory particle subunits, throughout the entire map. We were able to determine the architecture of the Rpn8/Rpn11 heterodimer, which had hitherto remained elusive.

The close relationship between HOMO-LUMO separations, symmetry considerations, and reactivity of the open shell in main group compounds is emphasized, as is their similarity in reactivity to transition metal organometallic compounds.\n\nThe unexpectedly potent reactivity of the heavier main group species arises U0126 from the large differences in bonding between the light and heavy elements. Specifically, the energy levels within the heavier element molecules are separated by much smaller gaps as a result of generally lower bond strengths. In addition, the ordering and symmetries of the energy levels are generally different for their light

counterparts. Such differences lie at the heart of the new reactions. Moreover, the reactivity of the Stem Cells & Wnt inhibitor molecules can often be interpreted qualitatively in terms of simple molecular orbital considerations. More quantitative explanations are accessible from increasingly sophisticated density functional theory (DFT) calculations.\n\nWe open with a short description of the background

developments that led to this work. These advances involved the synthesis and characterization of numerous new main group molecules involving multiple bonds or unsaturated configurations; they were pursued over the latter part of the last century and the beginning of the new one. The results firmly established that the structures and bonding in the new compounds differed markedly from those of their lighter element congeners.

The knowledge gained from this fundamental work provided the framework for an understanding of their structures and bonding, and hence an understanding of the reactivity of the compounds discussed here.”
“There is no doubt that distance is the principal parameter that sets the order of magnitude for electron-tunneling rates in proteins. However, there continue to be varying ways to measure electron-tunneling distances in proteins. This distance uncertainty blurs the issue of whether the intervening protein medium has been naturally selected to speed or slow any particular electron-tunneling reaction. For redox cofactors lacking metals, an edge of the GSI-IX supplier cofactor can be defined that approximates the extent in space that includes most of the wavefunction associated with its tunneling electron. Beyond this edge, the wavefunction tails off much more dramatically in space. The conjugated porphyrin ring seems a reasonable edge for the metal-free pheophytins and bacteriopheophytins of photosynthesis. For a metal containing redox cofactor such as heme, an appropriate cofactor edge is more ambiguous. Electron-tunneling distance may be measured from the conjugated heme macrocycle edge or from the metal, which can be up to 4.8 angstrom longer. In a typical protein medium, such a distance difference normally corresponds to a similar to 1000 fold decrease in tunneling rate.