This is attributed to two different homopolymerization reaction mechanisms, activated monomer (AM) and activated chain end APR-246 clinical trial (ACE), the former dominating at high temperature and the latter at low temperature. The effects of these homopolymerization reactions on the network and nanostructure of the nanocomposite are discussed, the important

aspect emerging being that the ACE mechanism is the one that most significantly enhances the exfoliation process. (C) 2009 Wiley Periodicals, Inc. J Appl Polym Sci 114: 1040-1047, 2009″
“We have studied copper phthalocyanine (CuPc) molecules deposited on a clean gold surface via synchrotron radiation photoemission. The persistent appearance of Au4f surface core-level emission in a series of depositions

suggests columnar growth of the adsorbed molecules covering only one-fifth of the surface. Molecular adsorption reduced the line intensity of emission from the Au surface slightly and also produced a shortened core-level shift, which demonstrates weakened s-d hybridization of the surface band. Moreover, manifestation of excess charge in the C1s core, along with an induced component in the Au4f core, indicates that charge polarization occurred at the organic/metal contact. The interface Vorinostat datasheet introduces an electronic state 1.09 eV below the Fermi level. Further analysis reveals that only the carbon atoms in the benzene rings are affected upon adsorption, suggesting that the molecular plane lies flat on the surface without changing the nominal bent configuration.”
“A

series of 0-3 composites of lead zirconate titanate (PZT) and nyton77 was prepared with PZT volume fractions between 0.1 and 0.6 using the combined method of solvent casting and hot pressing. Depending upon the volume fraction of PZT, the relative permittivities of the composites were found to be in the range of 200-2000 when measured at the highest practically possible temperature of 200 degrees C. As PZT volume fraction increases, both STA-9090 clinical trial the relative permittivity and the piezoelectric constant d(33) increase. The best combination of electrical properties and flexibility was obtained in 0.5 PZT volume fraction composites. At this composition, the remanent polarization P, of composites was up to 1000 mC/m 2 at the applied electric field of 40 MV/m and it increased with an increase in temperature. The highest piezoelectric strain coefficient d33 was obtained at 17 pC/N in 0.6 PZT volume fraction composites. (C) 2009 Wiley Periodicals, Inc. J Appl Polym Sci 114: 1048-1054, 2009″
“The local structure of a Zn, Sn codoped In(2)O(3) thin film grown on c-plane sapphire by pulsed-laser deposition was examined by polarization-dependent x-ray absorption spectroscopy.

FNS may help ameliorate mixed trauma spectrum syndromes. (The Journal of Neuropsychiatry and Clinical Neurosciences 2012; 24:237-240)”
“There are few published population-based data for psychogenic nonepileptic seizures (PNES). We prospectively

identified first presentations of PNES from a population of 367,566, over 3 years. PNES were diagnosed in 68 patients, in 54 of whom the diagnosis was confirmed by video/EEG recording, indicating an incidence of 4.90/100,000/year. Median diagnostic delay was 0.6 +/- 0.2 year. At presentation with PNES, our patients already had high rates of psychological morbidity, medically unexplained symptoms other than PNES, and economic dependence. At 3 months postdiagnosis, 27 of 54 patients (50.0%) were spell free. For 24 of the 27 patients (88.9%), spells GS-1101 mouse ceased immediately on communication of the diagnosis. At 6 months, 24 of 54 patients (44.4%) were spell free. Poor early outcome 3-deazaneplanocin A research buy was predicted by unemployment. Our data suggest that early outcome

is good in patients with recent-onset PNES, but some patients relapse quickly. (C) 2010 Elsevier Inc. All rights reserved.”
“CEA and CA19.9 are biomarkers routinely measured for monitoring treatment response in metastatic colorectal cancer (MCRC) patients, yet their predictive value during therapies containing new antineoplastic drugs (i.e. FOLFIRI/FOLFOX/Bevacizumab) has not yet been investigated.

Consecutive chemotherapy-naive MCRC patients treated with either standard chemotherapy-alone (FOLFIRI/FOLFOX) or chemotherapy+bevacizumab (FOLFIRI+bevacizumab) were included in the analysis. Patients had to have serial biweekly measurement of CEA and CA19.9 available for at least three months of treatment. Primary study endpoint was Progression PHA-739358 mw Free Survival (PFS). Biomarker levels and type of treatment as well as major demographic

and clinical factors were analyzed for their impact on PFS.

Out of 243 evaluated MCRC patients, 87 had biomarkers available as per inclusion criteria. Among all evaluated factors only type of treatment (chemotherapy-alone vs chemotherapy+bevacizumab) and baseline CA19.9 (> vs < normal) were independently associated with PFS, whilst neither baseline CEA nor biomarker reduction during therapy reached statistical significance. When patients with different baseline CA19.9 levels were analysed separately, only patients with abnormal CA19.9 benefited significantly from the administration of bevacizumab.

The current study demonstrated a significant predictive value of CA19.9, but not of CEA and biomarker reduction, for MCRC patients treated with new antineoplastic drugs. Moreover, only patients with abnormal baseline CA19.9 levels benefited significantly from bevacizumab.”
“Background: Malignant tumor cells may evoke the innate and adaptive immune systems.

The aim of this study is to investigate the role of ginsenoside-Rg1 in the microenvironment-dependent endothelial differentiation of human MSCs (hMSCs) in vitro. The endothelial differentiation environment was established by co-culturing hMSCs with mature endothelial cells (human umbilical vein endothelial cells) indirectly in vitro. Reverse transcriptase-polymerase chain reaction analysis and fluorescence immunocytochemistry showed a strong expression of endothelial-specific markers such as CD31, Von Willebrand factor, and VE-cadherin. Nepicastat Electron microscopy showed the endothelial characteristic Weibel-Palade bodies of differentiated hMSCs. The increased expression of CD31 demonstrated that Rg1

promoted the endothelial differentiation of hMSCs. The findings here show the differentiation of hMSCs into cells with phenotypic features of endothelial cells using indirect co-culture with mature endothelial cells and provide the evidence that ginsenoside-Rg1 can promote the milieu-dependent endothelial differentiation of hMSCs in vitro.”
“Objective: A “”review of reviews”" was undertaken to assess methodological issues in studies evaluating nondrug rehabilitation interventions in stroke patients.

Study Design and Setting: MEDLINE, ACY-241 Epigenetics inhibitor CINAHL, PsycINFO, and the Cochrane Database of Systematic Reviews were searched from January 2000 to January

2008 within the stroke rehabilitation setting. Electronic searches were supplemented by reviews of reference lists and citations identified by experts. Eligible studies were Crenolanib datasheet systematic reviews; excluded citations were narrative reviews or reviews of reviews.

Review characteristics and criteria for assessing methodological quality of primary studies within them were extracted.

Results: The search yielded 949 English-language citations. We included a final set of 38 systematic reviews. Cochrane reviews, which have a standardized methodology, were generally of higher methodological quality than non-Cochrane reviews. Most systematic reviews used standardized quality assessment criteria for primary studies, but not all were comprehensive. Reviews showed that primary studies had problems with randomization, allocation concealment, and blinding. Baseline comparability, adverse events, and cointervention or contamination were not consistently assessed. Blinding of patients and providers was often not feasible and was not evaluated as a source of bias.

Conclusions: The eligible systematic reviews identified important methodological flaws in the evaluated primary studies, suggesting the need for improvement of research methods and reporting. (C) 2012 Elsevier Inc. All rights reserved.”
“A 30-year-old manufacturing process for the biologic product l-asparaginase from the plant pathogen Erwinia chrysanthemi was rigorously qualified and validated, with a high level of agreement between validation data and the 6-year process database.

Reduction estimates were similar among infants and those aged 1-4 years of age, even though only 25% of the latter group was likely to have received vaccine by early 2008. Estimated coverage with >= 1 dose of

rotavirus vaccine among infants increased from 63% at the end of 2006 to 94% ASP2215 mw at the end of 2008.

Conclusions: RV1 appears to have had a substantial impact on diarrhea-associated hospitalizations among young children in Panama.”
“Background and Aims: Increased platelet aggregation is a major risk factor for heart attacks, stroke and thrombosis. Long chain omega-3 polyunsaturated fatty acids (LCn-3PUFA; eicosapentaenoic acid, EPA; docosahexaenoic acid, DHA) reduce platelet Selleckchem BMS-754807 aggregation; however studies in the published literature involving EPA and/or DHA supplementation have yielded equivocal results. Recent in vitro studies have demonstrated that inhibition of platelet aggregation by LCn-3PUFA is gender specific. We examined the acute effects of dietary supplementation with EPA or DHA rich oils on platelet aggregation in healthy male and females.

Methods and Results: A blinded placebo controlled trial involving 15 male and 15 female subjects. Platelet aggregation was measured at 0, 2, 5 and 24 h post-supplementation with a single dose of either a placebo or EPA or DHA rich oil capsules. The relationship between

LCn-3PUFA and platelet

activity at each time point was examined according to gender vs. treatment. EPA was significantly the most effective in reducing platelet aggregation in males at 2, 5 and 24 h post-supplementation (-11%, -10.6%, -20.5% respectively) whereas DHA was not effective relative to placebo. In contrast, Selleckchem SIS3 in females, DHA significantly reduced platelet aggregation at 24 h (-13.7%) while EPA was not effective. An inverse relationship between testosterone levels and platelet aggregation following EPA supplementation was observed.

Conclusion: Interactions between sex hormones and omega-3 fatty acids exist to differentially reduce platelet aggregation. For healthy individuals, males may benefit more from EPA supplementation while females are more responsive to DHA. (C) 2010 Elsevier B.V. All rights reserved.”
“Study Design. Retrospective literature review.

Objective. To assess the quality of randomized controlled trial reporting in spine surgery.

Summary of Background Data. The use of the Consolidated Standards of Reporting Trials (CONSORT) criteria in clinical trials aims to improve adherence to a set of generally accepted practices in the reporting of randomized controlled trials. Randomized controlled trials in spine surgery are important sources of evidence-based practice, but the quality of their execution and reporting have not been reported adequately.

Methods.

Although there are beneficial nutritional properties, the prevailing mycobiota of Brazil

nuts include fungi that are producers of aflatoxins, such as Aspergillus flavus. A. parasiticus and A. nomius. Aflatoxins have deleterious effects in consumption considering the global distribution chain, affecting major exporting countries. The present review is focused on the importance of Brazil nuts for the Amazon rainforest, emphasizing on the social and environmental impact of its production, on the mycobiota contamination of seeds, and on the presence of mycotoxins and related food safety aspects. (C) 2011 Elsevier Ltd. All selleck chemical rights reserved.”
“Colorectal cancer presents as an emergency with large bowel obstruction in up to 29% of cases. These patients are often elderly with multiple co-morbidities and deranged physiological function. Traditionally, the mainstay of treatment was with staged surgical procedures, but recent trends have moved towards a primary resection and anastomosis. The use of self-expanding metallic stents has been described to alleviate obstruction and act as a bridge to surgery or as a definitive palliative procedure.

Although there is no significant difference in long-term survival between these two treatment modalities, there is no consensus as to which approach is optimal in this group patients. (C) 2010 The Royal College of Radiologists. Published by Elsevier Ltd. All rights reserved.”
“Eriocheir sinensis is a euryhaline crab migrating from sea to freshwater habitats during the juvenile THZ1 in vivo stage. We used quantitative

real-time polymerase chain reaction (qRT-PCR) to investigate the gene expression profile of Na+-K+-ATPase, Hsp70 (heat shock protein 70) and Hsp90 in megalopa exposed to salinities of 0, 2, 5, 10, and 15 parts per thousand. Both low and high salinities markedly stimulated expression of Na+-K+-ATPase, Hsp70 and Hsp90 genes of Chinese mitten crab megalopa; salinity had different effects on Na+-K+-ATPase, Hsp70 and Hsp90 levels depending on the duration of salinity stress, implying that Na+-K+-ATPase, Hsp70 and Hsp90 may play an important role in salinity tolerance in this crab species.”
“Protein-polysaccharide interactions play an essential structure-controlling role in foods and biomaterials. Turbidity and Selleck Navitoclax electrophoretic mobility measurements were used to investigate the formation of soluble and insoluble complexes between pea protein isolate (PPI) and the cationic polysaccharide. chitosan (Ch) as a function of pH and biopolymer mixing ratio (1:1-20:1 PPI-Ch). In addition, pH-induced conformational changes of PPI upon complexation with Ch were studied by fluorometry. As the PPI-Ch mixing ratio increased from 1:1 to 12.5:1, critical structure forming events (i.e., those associated with the formation of soluble and insoluble complexes) shifted to higher pHs, and progressively behaved similar to those for PPI alone.

Methods: Four adult patients with refractory OCD were implanted with Model 3387 leads bilaterally in an area of VC/VS. Postoperative test stimulation was performed at least 2 weeks after surgery. We performed double-blinded postoperative test stimulation with different contact and voltage. The relationship of stimulation-induced smile/laughter and chronic response was examined.

Results: Patients presented smile, laughter, euphoria, increased heart rate, increased blood pressure, smell, chest vibration, dizziness, nausea, heat, or increased sexual drive during acute stimulation. We found that the higher the percentage Selleck MK2206 of smile/laughter

(34.3%, 31.3%, 56.3%, and 12.5% for four cases), the greater SBE-β-CD in vitro the reduction in the Yale-Brown Obsessive Compulsive Scale (30.6%, 38.9%, 58.8%, and 7.7% respectively at 15-month DBS).

Conclusion: This study showed that acute DBS of the VC/VS might cause mood change, cardiovascular, sensory, or motor effects. These effects were transient or habituated over six months. We suggest stimulation-induced smile/laughter may be a possible predictor for long-term DBS outcome. Larger studies, genetic studies, and imaging studies are needed to evaluate the effects of different parameters and possible predictors in the treatment of OCD.”
“Early and effective

treatment of chronic inflammatory demyelinating polyneuropathy (CIDP) is important to minimize axonal degeneration that occurs secondary to demyelination. The disease course is invariably chronic, so long-term treatment is often required, and adverse effects and costs are important

considerations in devising a treatment plan. CIDP responds to prednisone, but long-term treatment can result in significant adverse effects. Azathioprine, mycophenolate mofetil, and cyclosporine can be used as steroid-sparing Crenolanib solubility dmso agents and may facilitate more rapid and successful tapering of prednisone. Intravenous immunoglobulin (IVIg) and plasma exchange are also effective in the treatment of CIDP and can be used in patients who are unresponsive to prednisone or develop steroid-related adverse effects. IVIg may also be used as a first-line treatment, but its cost can be a limiting factor. A few uncontrolled studies have suggested that pulsed weekly methylprednisolone is both effective and well tolerated in the long-term treatment of CIDP. Treatments based on rituximab or cyclophosphamide have also been used in resistant disease.

Variants of CIDP have been described on the basis of their association with specific antibodies or immunoglobulins and their response to specific immunomodulatory treatments. Multifocal motor neuropathy with conduction block responds to IVIg in the majority of patients. However, weakness may slowly worsen over time, and some patients become unresponsive.

67, P = 8.6 x 10(-5)).

Conclusions: SLC30A8 SNPs and haplotypes are associated with GADA negative diabetes in Malaysian subjects, and this association is markedly higher among Malaysian Indian subjects.”
“Background and Purpose: Endourology has evolved rapidly for the management of both benign and malignant disease of the upper urinary tract. Limited data exist, however, on the occupational hazards posed by complex endourologic procedures. The aim of this study was to determine the prevalence and possible causes of hand

problems among endourologists Elafibranor supplier who routinely perform flexible ureteroscopy compared with controls.

Materials and Methods: An online computer survey targeted members of the Endourological Society and psychiatrists in academic and community settings. A total of 600 endourologists and 578 psychiatrists were contacted by e-mail. Invited physicians were queried regarding their practice settings and symptoms of hand pain, neuropathy, and/or discomfort.

Results: Survey responses were obtained from 122 (20.3%) endourologists and 74 (12.8%) psychiatrists. Of endourologists, 61% were in an academic setting and 70% devoted their practice to endourology. Endourologists were in practice for a mean 13 years, performing 4.5 ureteroscopic cases per week with a mean operative time of 50 minutes. Hand/wrist problems were reported by 39 (32%) endourologists compared

with 14 (19%) psychiatrists (P = 0.0486, Staurosporine manufacturer relative risk [RR] = 1.69). Surgeons who preferred counterintuitive ureteroscope deflection were significantly more likely to have problems (56%) compared with Selleckchem CDK inhibitor intuitive users (27%) (RR 2.07, P = 0.0139) or those with no preference

(26%) (RR 2.15, P = 0.0451). Overall, most respondents (85%) with hand/wrist problems needed either medical or surgical intervention.

Conclusions: Hand and wrist problems are very common among endourologists. Future studies are needed to develop more ergonomic platforms and thereby reduce the endourologist’s exposure to these occupational hazards.”
“We compared rifabutin susceptibility and rpoB mutations in 98 multi-drug-resistant strains of Mycobacterium tuberculosis (MDR-TB) by DNA sequencing and with a line probe assay using the commercially available INNO-LiPA Rif. TB kit (the LiPA). Our results indicated that rifabutin continues to remain active against MDR-TB strains harboring certain genetic alterations and also that the LiPA might be useful in identifying MDR-TB strains susceptible to rifabutin.”
“Pectic polysaccharides have attracted great attention due to their health-promoting potential. From the Styrian oil-pumpkin biomass representing an alternative pectin source, two series of acidic polysaccharide fractions were isolated using in succession hot water. EDTA and dilute HCl. Chemical and spectroscopic (FTIR.

This lily appears to be poorly synchronized with its pollinators. Across the years of the study, pollination limitation appears to be increasing, perhaps because the synchronization is getting worse.”
“Background: Increased control has produced remarkable reductions of malaria in some parts of sub-Saharan Africa, including Rwanda. In the southern highlands, Buparlisib manufacturer near the district capital of Butare (altitude, 1,768 m), a combined community-and facility-based survey on Plasmodium infection was conducted early in 2010.

Methods: A total of 749 children below five years of age were examined including 545 randomly

selected from 24 villages, 103 attending the health centre in charge, and 101 at the referral district hospital. Clinical, parasitological, haematological, and socio-economic data were collected.

Results: Plasmodium falciparum infection (mean multiplicity, 2.08) was identified by microscopy and PCR in 11.7% and 16.7%, respectively; 5.5% of the children had malaria. PCR-based P. falciparum prevalence ranged between 0 and 38.5% in the villages, and was 21.4% in the health centre, and 14.9% in the hospital. Independent predictors of infection included increasing age, low mid-upper arm circumference, absence of several household

assets, reported recent intake of artemether-lumefantrine, and chloroquine in plasma, measured by ELISA. Self-reported bed net use (58%) reduced Stem Cell Compound Library chemical structure infection only in univariate analysis. In the communities, most infections were seemingly asymptomatic but GW4869 anaemia was observed in 82% and 28% of children with and without

parasitaemia, respectively, the effect increasing with parasite density, and significant also for submicroscopic infections.

Conclusions: Plasmodium falciparum infection in the highlands surrounding Butare, Rwanda, is seen in one out of six children under five years of age. The abundance of seemingly asymptomatic infections in the community forms a reservoir for transmission in this epidemic-prone area. Risk factors suggestive of low socio-economic status and insufficient effectiveness of self-reported bed net use refer to areas of improvable intervention.”
“Background: In our practice, we noticed that patients respond differently when pacemaker insertion is recommended to them. We then saw the need to study how African patients accept elective pacemaker implantation.

Aim: The study was conducted to determine African patients’ responses to elective pacemaker insertion.

Patients and Methods: We recruited patients who reported at our outpatient clinic with indications for elective pacemaker insertion. The study period lasted 64 months from September 1999. The patients were counseled before insertion and we sought their consent before being recruited for the study. Five possible responses were used to grade acceptance. They were followed up for 12 months each.

The aim of the present study was to investigate the differential activation of brain regions related to the reward circuitry after acquistion/expression of cocaine CPP or social interaction CPP. Our findings indicate that cocaine CPP and social interaction CPP activated almost the same brain regions. However, the granular insular cortex and the dorsal part of the granular insular cortex were more activated after cocaine CPP, whereas the prelimbic cortex and the core subregion of the nucleus accumbens were more activated after social interaction

FK228 ic50 CPP. These results suggest that the insular cortex appears to be potently activated after drug conditioning leraning while activation of the prelimbic cortex-nucleus accumbens core

projection seems to be preferentially involved in the conditioning to non-drug stimuli such as social interaction.”
“Increased concentrations of circulating insulin-like growth factor-I (IGF-I) or IGF-I relative to IGF-binding proteins (IGFBPs) are associated with increased risk of developing several forms of cancer. Conversely, exercise is linked with reduced risk. This study aims to investigate the effect of a low-intensity exercise program on circulating levels of IGF-I, IGFBP-1, and IGFBP-3, in previously sedentary males. Fourteen click here healthy men participated in cycle ergometer training at lactate threshold intensity for 60 min/day,

5 days/week for 6 weeks. After aerobic training, insulin sensitivity improved by 20%, while fasting insulin levels decreased by 13%. Simultaneously, low-intensity aerobic training decreased the circulating levels of IGF-I by 9%, while IGFBP-1 levels increased by 16%. An interesting finding was that higher pretraining level of IGF-I was associated with greater decline in IGF-I with training. Insulin-sensitizing low-intensity Gamma-secretase inhibitor aerobic exercise is thus considered to be an effective method for downregulating IGF-I and upregulating IGFBP-1 levels.”
“On earth, gravity vector conditions the development of all living beings by physically imposing an axis along which to build their organism. Thus, during their whole life, they have to fight against this force not only to maintain their architectural organization but also to coordinate the communication between organs and keep their physiology in a balanced steady-state. In space, astronauts show physiological, psychological, and cognitive deregulations, ranging from bone decalcification or decrease of musculature, to depressive-like disorders, and spatial disorientation. Nonetheless, they are confronted to a great amount of physical changes in their environment such as solar radiations, loss of light-dark cycle, lack of spatial landmarks, confinement, and obviously a dramatic decrease of gravity force.

Secondary analyses included incidence of adverse events (ITT) and treatment adherence rate (completers).

Results: In total, 1250 patients comprised the ITT population: 32.6% with newly diagnosed BMS-777607 hypertension, 40.5% with hypertension uncontrolled

with monotherapy; and 26.9% with hypertension inadequately managed with another combination therapy. Mean SBP/DBP decreased significantly from baseline (167.4 +/- 15.2/101.4 +/- 9.1 mmHg) over 60 days (-41.9 +/- 34.8/-23.2 +/- 21.8 mmHg; p < 0.0001). Target BP was achieved in 66.1% of patients in the total population, 68.3% of untreated patients, 68.4% of patients uncontrolled with monotherapy, and 59.9% of patients inadequately managed with combination therapy. In 161 patients with SBP > 180 mmHg at baseline

(newly diagnosed: n = 50; uncontrolled on monotherapy: n = 53; inadequately managed on combination therapy: n = 58), BP was reduced by 63.2 +/- 32.5/29.0 +/- 21.9 mmHg (p < 0.0001) at day 60. The fixed combination was safe and well tolerated. All 1175 patients completing the buy Alisertib 60-day study (94%)adhered to their treatment regimen.

Conclusion: Fixed combination perindopril/amlodipine was found to be an effective and well tolerated antihypertensive treatment, with an excellent rate of treatment adherence in the clinical setting. Fixed combination perindopril/amlodipine is expected to be useful in the management of hypertension in primary healthcare, with a positive

impact www.selleckchem.com/products/qnz-evp4593.html on treatment adherence.”
“In this study, a novel well-defined epoxy mid-chain functional macromonomer of poly(e-caprolactone) (PCL) has been synthesized by ring-opening polymerization (ROP) of e-caprolactone (e-CL) and epoxidation on workup with 3-chloroperoxybenzoic acid. The ROP of e-CL monomer in bulk at 110 degrees C, by means of a dihydroxy functional initiator namely, 3-cyclohexene-1,1-dimethanol in conjunction with stannous-2-ethylhexanoate, (Sn(Oct)2), yielded a well-defined PCL with a cyclohexene mid-chain group. The epoxidation of the cyclohexene (CH) mid-chain group of PCL was performed using 3-chloroperoxybenzoic acid. GPC, IR, and 1H-NMR analyses revealed that a low-polydispersity macromonomer of PCL with the desired cyclohexene oxide (CHO) functionality at the mid-chain was obtained. The photoinduced cationic polymerizations of this macromonomer yielded comb-shaped and graft copolymers. (c) 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012″
“Study Design. Secondary analysis within a large clinical trial.

Objective. To evaluate the changes in treatment preference before and after watching a video decision aid as part of an informed consent process.

Summary of Background Data. A randomized trial with a similar decision aid in herniated disc patients had shown decreased rate of surgery in the video group, but the effect of the video on expressed preferences is not known.

Methods.