These data present guidance to clinicians and other professionals investigating the deficiency of essential trace metals in biological samples (scalp hair and blood) of children with night blindness.”
“The meropenem susceptibility surveillance
study was performed between 2002 and 2006 in Japan. Meropenem resistance against Pseudomonas aeruginosa was nearly constant at about 10% during the period. For all species of the bacteria tested, the MIC90 of meropenem in 2006 was not more than 4-fold higher than that in 2002 and 2004. (C) 2008 Elsevier Inc. All rights reserved.”
“The years 2000 through mid-2010 marked a transformational period in understanding of the biosynthesis of marine natural products. During this decade the field emerged from one largely dominated by chemical approaches to understanding biosynthetic pathways to one selleck products incorporating the full force of modern molecular biology and bioinformatics. Fusion of chemical and biological approaches yielded great advances in understanding the genetic and enzymatic basis for marine natural product biosynthesis. Progress was particularly pronounced for marine microbes,
especially actinomycetes and cyanobacteria. During this single decade, both the first complete marine microbial natural product biosynthetic gene cluster sequence was released as well as the first entire genome sequence for a secondary metabolite-rich marinemicrobe. The decade also saw tremendous progress CHIR-99021 inhibitor in recognizing the key role of marine microbial symbionts of invertebrates in natural product biosynthesis. Application of genetic and enzymatic knowledge led to genetic engineering of novel “unnatural” natural products during this time, as well as opportunities for discovery of novel natural products through genome mining. The current review highlights selected seminal studies from 2000 selleck screening library through to June 2010 that illustrate breakthroughs in understanding of marine natural product biosynthesis
at the genetic, enzymatic, and small-molecule natural product levels. A total of 154 references are cited.”
“The interaction between a common soil yeast, Cryptococcus laurentii, and a slow-growing medicinal plant adapted to low-nutrient soils, Agathosma betulina (Berg.) Pillans, was studied. C. laurentii CAB 578 was isolated from the rhizosphere of wild A. betulina, and liquid chromatography-tandem mass spectrometry (LC-MS-MS) analysis revealed that the yeast was capable of producing polyamines, such as cadaverine and spermine, while growing in vitro in a chemically defined medium. Since the exogenous application of polyamines are known to impact on root growth, these findings supported the results obtained when axenic cultures of A. betulina seedlings were inoculated with C. laurentii CAB 578 and cultivated for 5 months under glasshouse conditions. The presence of the yeast increased root growth by 51%.
These observations support the conclusion that, during the nuclear cataract formation, alterations in protein packing are extensive and can result in pronounced density fluctuations. Aging causes the MLB cores to become increasingly different in their protein packing from the adjacent cytoplasm. These results support the hypothesis that the MLBs increase their scattering with age and nuclear cataract formation. (C) 2012 Elsevier Ltd. All rights reserved.”
“What is known and Objective: Invasive fungal infections are a major threat
to renal transplant recipients. Micafungin and voriconazole are two useful antifungal agents for treating such infections. Our objective is to evaluate the comparative efficacy and safety of micafungin and voriconazole in the initial treatment of such infections.\n\nMethods: Selleck AZD9291 In this prospective, multicentre, open-labelled, randomized, controlled trial, renal transplant recipients with invasive fungal infections were assigned to receive either micafungin or voriconazole. The enrolled
subjects received a kidney transplant between March 2008 and March 2010 at one of the two transplant centres in Henan Province, China. The efficacy and adverse effects of the two treatments were compared.\n\nResults and Discussion: The clinical trial enrolled 65 patients, of whom 31 were treated with micafungin, and 34 with voriconazole. The rates of microbiological evidence of infection in the micafungin and voriconazole groups were 64.5% and 70.5%, respectively, whereas the rates of Candida as the major cultured fungus were 80.0% and 75.0%, respectively. Complicated bacterial infection rates in the BKM120 mw two treatment groups were 38.7% and 32.4%, respectively, whereas complicated CMV viral infection occurred at a rate of 19.2% and 23.5%, respectively. Fungal infection within one to 3 months after transplant was 83.6% (26/31) and 85.3% (29/34) in the micafungin and voriconazole groups, respectively. There was no significant difference between the two groups G418 cost in terms of efficacy, survival beyond 10 days
and discontinuation of treatment because of lack of efficacy (P > 0.05). Mortality rates in the micafungin and voriconazole groups were 9.7% (3/31) and 12.1% (4/33), respectively. Rates of adverse effects in the two groups were 41.9% and 51.6% (P > 0.05), respectively.\n\nWhat is new and Conclusions: This is the first comparison of micafungin and voriconazole in renal transplant patients. Our study shows that the effectiveness of micafungin was similar to that of voriconazole in such patients.”
“Aims: To compare the effects of lifestyle modification programs that prescribe low-glycemic load (GL) vs. low-fat diets in a randomized trial.\n\nMethods: Seventy-nine obese adults with type 2 diabetes received low-fat or low-GL dietary instruction, delivered in 40-week lifestyle modification programs with identical goals for calorie intake and physical activity.
However, the precise https://www.selleckchem.com/products/epoxomicin-bu-4061t.html role of resistance to AZD6244 is largely unknown. We and other groups have reported that the novel oncogene Metadherin (MTDH) is associated with multiple drug resistance, but there is no report about its role in treatment of AZD6244. Here we report that the resistance to AZD6244 can be reserved by downregulating MTDH in breast cancer cell lines. When the MTDH was downregulated, the breast cancer cells exhibited a significantly increased sensitivity to AZD6244 as measured by MTT assay. After treated with AZD6244 the MTDH-knockdown cells showed more apoptosis rate and growth inhibition.
We also showed that knockdown of MTDH cannot only increase expression of FOXO3a but also activate it by promoting its translocation via MTDH/ERK1/2/FOXO3a pathway rather than MTDH/AKT/FOXO3a pathway. In conclusion knockdown MTDH can enhance the breast cancer cells sensitivity to AZD6244 via regulating the expression and activity of FOXO3a. These indicate us that MTDH is a candidate marker to predict the clinical efficacy of AZD6244 and targeting MTDH could overcome the resistance to AZD6244 in breast cancer cells.”
the relationship between diabetes and impairments in functional and cognitive status as well as depression in older people.\n\nCross-sectional study.\n\nElderly Health Centres (EHC) in Hong Kong.\n\n66,813 older people receiving baseline assessment at EHC in 1998 to 2001.\n\nDiabetes status was defined CA4P in vivo by self-report and blood glucose tests. Functional status was assessed by 5 items of instrumental activities of daily living (IADL) and 7 items of activities of daily living (ADL). Cognitive status was screened by the Abbreviated Mental Test-Hong Kong version (AMT). Depressive symptoms were screened by the Geriatric Depression Scale-Chinese version (GDS).\n\nAmong
the subjects, 10.4% reported having regular treatment for diabetes, 3.4% had Kinase Inhibitor Library molecular weight diabetes but were not receiving regular treatment, and 86.2% did not have diabetes. After controlling for age, sex and education level, those having regular treatment for diabetes were 1.7 times more likely (OR=1.65, 95% CI: 1.51-1.80) to have functional impairment, 1.3 times more likely (OR=1.28, 95% CI: 1.11-1.48) to have cognitive impairment and 1.3 times more likely (OR=1.35, 95% CI: 1.25-1.46) to have depression, than older people without diabetes.\n\nOlder people with diabetes may be less capable of managing the disease than the younger ones as a result of increased risk of both physical and cognitive impairment. This study provided further evidence for the need of an international consensus statement regarding care of diabetes in older people.”
“Canna edulis ker is a potential feedstock for ethanol production because of its low nutrition requirements and the high starch content of its tubers. The processing of C. edulis is limited by the high viscosity of the biomass.
01). Some embedded microcalcifications in the tomosynthesis phantoms were visible only in the scatter-corrected reconstructions. The visibility of the findings in two patient images was also improved by the application of the scatter correction algorithm. The MTF of the images did not change after application of the scatter correction algorithm, indicating that
spatial resolution was not adversely affected.\n\nConclusions: Our software-based scatter correction algorithm exhibits great potential in improving the image quality of DBT acquisitions of both phantoms and patients. The proposed algorithm does not require a time-consuming MC simulation for each specific case to Sapanisertib be corrected, making it applicable in the clinical realm. (C) 2011 American Association of Physicists in Medicine. [DOI: 10.1118/1.3659703]“
“Aging hearts are known to have diminished capacity PD173074 price to be protected
against reoxygenation ischemia/reperfusion (IR) injury provided by various cardioprotective regimens. In search of a more successful regimen, we have studied the response of aged hearts to preconditioning (PC) and postconditioning (POST) elicited by sphingosine or sphingosine 1-phosphate treatment. An ex vivo rat heart model was used to study the ability of PC and POST to protect old hearts (27 month) against I/R injury generated by 40 minutes (min) of index ischemia followed by 40 min of reperfusion. The response to ischemic PC was reduced in 27 month old hearts relative to
3-6 month (young) hearts as noted by a poor recovery of left ventricular developed pressure (LVDP) upon reperfusion (45% vs. 74% in young hearts) and a large infarct size after 40 min of reperfusion (37% versus 8% in young hearts). PC with sphingosine I-phosphate (SIP) was also poor in old hearts yielding only 49% recovery of LVDP and a 27% infarct size. In contrast, PC with sphingosine was unaffected by aging; the 78% recovery of LVDP and 8% infarct size were not different from young hearts. Ischemic POST was less affected by aging than ischemic PC, but the old hearts still experienced infarct sizes of 28%. POST of old hearts with SIP was also associated with Kinase Inhibitor Library concentration a substantial infarct size (24%). However, POST of old hearts with sphingosine was superior to the other forms of POST in that it reduced the infarct size to 12%. SIP levels were found to be lower in old hearts which may contribute to the decreased effectiveness of ischemic PC and POST. Further, phospho-Akt levels and distribution were altered in response to cardioprotection in the old hearts. In conclusion, POST was less affected by aging than PC; and sphingosine is a uniquely effective agent for both PC and POST of aging hearts.”
“Objective: To assess the association between recent cigarette smoking (CS) in female and male partners and assisted reproduction technology (ART) outcomes.\n\nDesign: Cohort prospective study.\n\nSetting: University ART program in Chile.
A two-cuff technique was used for orthotopic liver transplantation, and HOCs were subsequently transplanted into recipients. Biochemical indicators of liver function were assessed 4 weeks after transplantation. HOC transplantation significantly prolonged the median survival
time and improved the liver function of rats receiving HOCs compared to controls (P = 0.003, Student t-test). Administration of HOCs to rats also receiving liver transplantation significantly reduced acute allograft rejection compared to control liver transplant rats 3 weeks following transplantation (rejection activity index score: control = 6.3 +/- 0.9; HOC = 3.5 +/- 1.5; P = 0.005). These results indicate that HOCs may be useful in therapeutic liver regeneration after orthotopic liver transplantation.”
“Highly learn more stable dispersions of nanosized silver particles were synthesized using a straightforward, cost-effective, and ecofriendly method. Nontoxic glucose was utilized as a reducing agent and poly-alpha, AZD1480 cost gamma, L-glutamic acid (PGA), a naturally occurring anionic polymer, was used as a capping agent to protect the silver nanoparticles from agglomeration and render them biocompatible. Use of ammonia during synthesis was avoided. Our study clearly demonstrates how the concentration of the capping agent plays a major role in determining the dimensions,
morphology, and stability, as well as toxicity of a silver colloidal solution. Hence, proper optimization is necessary to develop silver colloids of narrow size distribution. The samples were characterized by Fourier transform infrared spectroscopy, ultraviolet-visible spectroscopy, ACY-241 inhibitor field-emission scanning electron microscopy, transmission electron microscopy, and zeta potential measurement. MTT assay results indicated good biocompatibility of the PGA-capped silver nanoparticles. Formation of intracellular reactive oxygen species was measured spectrophotometrically using 2,7-dichlorofluorescein diacetate as a fluorescent probe, and it was shown that the PGA-capped silver nanoparticles did not induce intracellular formation of reactive oxygen species.”
“The Practice Advisory on Liposuction
published by the American Society of Plastic Surgeons provides a thorough review of anesthetic techniques and guidelines for surgeons who perform liposuction. However, there is evidence to support several changes to the anesthetic infiltrate guidelines that will improve patient safety. These proposed recommendations will have the most impact on patients undergoing office-based procedures, where dedicated anesthesia providers may not be present, but they should also guide practice in both ambulatory care centers and hospitals. The primary foci of the proposed changes include restrictions on bupivacaine use and creation of lidocaine concentration guidelines.”
“A fluorescent assay for the evaluation of inhibitors of fatty acid amide hydrolase (FAAH) is described.
Critical twinning stress of cadmium zinc telluride (CdZnTe or CZT) calculated is 1.38 GPa. All the twin boundaries are along the (11-1) orientation, except the one with the (-111) plane that supports
the indentation, interpreting the unidirectional and boundary-free characteristics, confirmed in the experiment. Three twin thicknesses after unloading are 3.2, 3.5, and 16 nm, which is consistent with the experimentally repeated this website pattern of a lamellar twin with thickness larger than 12.7 nm, followed by one or several twins with thicknesses smaller than 12.7 nm. An inverse triangle of a twin combining with three twins generate a synergistic strengthening effect through the hardening and softening functions, illuminating the ultrahigh hardness demonstrated in the experiment. Twinning takes place in loading, and detwinning occurs in unloading, which expounds the high ductility observed in the experiment.”
“Phosphatidylethanolamine (PE) is an important inner membrane phospholipid
mostly synthesized de novo via the PE-Kennedy pathway and by the decarboxylation of phosphatidylserine. CTP:phosphoethanolamine cytidylyltransferase (Pcyt2) catalyzes AZD7762 chemical structure the formation of CDP-ethanolamine, which is often the rate regulatory step in the PE-Kennedy pathway. In the current investigation, we show that the reduced CDP-ethanolamine formation in Pcyt2(+/-) mice limits the rate of PE synthesis and increases the availability of diacylglycerol. This results in the increased formation of triglycerides, which is facilitated by stimulated de novo fatty acid synthesis and increased uptake of pre-existing fatty acids. Pcyt2(+/-) mice progressively accumulate more diacylglycerol and triglycerides with age and have modified fatty acid composition, predominantly in PE and triglycerides. Pcyt2(+/-) additionally
have an inherent blockage in fatty acid utilization as energy substrate and develop impaired tolerance to glucose and insulin at an older age. Accordingly, Stattic in vivo gene expression analyses demonstrated the up-regulation of the main lipogenic genes and down-regulation of mitochondrial fatty acid beta-oxidation genes. These data demonstrate for the first time that to preserve membrane PE phospholipids, Pcyt2 deficiency generates compensatory changes in triglyceride and energy substrate metabolism, resulting in a progressive development of liver steatosis, hypertriglyceridemia, obesity, and insulin resistance, the main features of the metabolic syndrome.”
“In 1999, the American Academy of Pediatrics (AAP) issued a policy statement addressing media use in children. The purpose of that statement was to educate parents about the effects that media-both the amount and the content-may have on children.
In conclusion, when followed correctly, this modified Selleckchem MK-2206 cryoloop
vitrification protocol had little effect on the survival rate and development potential of mature mouse oocytes.”
“The abundance of marine benthic organisms often exhibits distinct distributional patterns, which is generally governed by many physical and biological factors specific to the habitat. In this study, the spatial and temporal variations in abundance of the dog conch, Strombus canarium Linnaeus 1758, a commercially important marine gastropod, was investigated. Assessment of conch abundance at Merambong seagrass bed, Malaysia, was conducted using a transect belt method. Sampling stations were randomly selected and environmental parameters associated with the habitat were recorded. The species find more showed distinct spatial distributional pattern. Conch densities were significantly higher in sheltered areas, mainly in mixed seagrass bed dominated by Halophila spp. and with high sediment organic content. The densities were relatively very low in areas dominated by the tape seagrass, Enhalus acoroides. The species studied also showed distinct temporal variation in abundance. The abundance value was seasonally varied with highest density recorded
during the wet monsoon season (p smaller than 0.05). The densities were otherwise very low during the dry season, except for a slight peak in July. Since the conch is a very important fishery species within the Johor Straits and regulations on their harvesting is still lacking, this information ASP2215 molecular weight would be very important for their sustainable management.”
“This study is a nationwide survey of all clinical methicillin-resistant Staphylococcus aureus (MRSA) isolates, including community-acquired MRSA (CA-MRSA), in Japan. A total of 857 MRSA clinical isolates were collected from
the 16 institutions throughout Japan that participated in the survey (2008-2009). The drug susceptibility and staphylococcal cassette chromosome mec (SCCmec) typing and the presence of specific pathogenic genes were evaluated. The isolates comprised SCCmec type II (73.6%), type IV (20%), and type I (6%). The percentage of SCCmec type IV isolates was significantly higher in outpatients than in inpatients. Most of the isolated strains were sensitive to vancomycin (VCM, MIC <= 2 mu g/mL), linezolid (MIC <= 4 mu g/mL), and teicoplanin (MIC <= 8 mu g/mL). Although most strains were sensitive to VCM, the MIC value of VCM for SCCmec type II strains was higher than that for SCCmec type IV strains. Only 4 (2.3%) of 171 SCCmec type IV strains were Panton-Valentine leukocidin (lukS/F-PV)-positive.
PRP with insulin synergistically upregulated fibroblast growth factor receptor (FGFR) and downregulated epidermal growth factor receptor (ErbB) expression; moreover, PRP in association
prevented insulin-induced insulin-like growth factor-1 receptor and insulin receptor downregulation. The inhibition of FGFR-1, epidermal growth factor receptor (EGFR), and epidermal growth factor receptor-2 (ErbB2) activity reduced ASC proliferation, but only that of FGFR-1 reduced adipogenesis and Akt phosphorylation, selleck chemicals llc whereas the ErbB2 inhibition effects were the opposite. However, EGFR activity was needed for ErbB2-mediated inhibition of ASC adipogenesis. Clinically, the injection of insulin further ameliorated patients’ 1-year PRP-induced fat graft volume maintenance and contour restoring. Our results ascertain that PRP in association with insulin greatly potentiates adipogenesis in human ASCs through a FGFR-1 and ErbB2-regulated Akt mechanism. The ameliorated clinical fat graft maintenance suggests additional useful translational applications of combined PRP-insulin treatment in regenerative medicine. STEM CELLS TRANSLATIONAL MEDICINE 2012;1:206-220″
“The label free real time surface plasmon resonance based immuno-sensing system was first
time demonstrated in the authors’ lab using a mercaptoundecanoic acid modified gold sensor chip. The kinetics of the antigen-antibody interaction was determined for optimization of resonance signal intensity PX-478 for the detection of antigen(s) of fungal teliospores-causative entities of Karnal bunt disease of wheat incited by Tilletia indica. The approach involves the use of a rabbit polyclonal (anti-teliospore) antibody and mono-specific antibody generated against intact teliospores and purified diagnostic antigen of 28 kDa protein. The sensitivity of each of these two immunosensors constructed with the different concentrations of teliosporic antigen was determined by sensogram analysis
which showed detection GDC-0994 sensitivity as low as 625 and 156 pg which is equivalent to 2.5 and 1.0 teliospores over the sensor surface by getting an angle of dip indicating greater affinity of raised anti-teliospore antibodies and monospecific antibody respectively. By using kinetic evaluation software, the equilibrium constant (KD) and maximum binding capacity of analyte (B-max) values were calculated and found to be 3.45 and 331.8 nM respectively using polyclonal and 1.57 and 289.57 nM respectively using monospecific antibody.”
“An in vitro system was used to analyze the effects of sex steroids on the development of primary (late perinucleolar stage) and early secondary, previtellogenic (early cortical alveolus stage) ovarian follicles of coho salmon cultured for up to 21 days.
Thirty-nine additional blind-coded Bartonella strains were correctly identified at the species level,
including 36 with a significant score. Altogether, these data demonstrate that MS is an accurate and reproducible tool for rapid learn more and inexpensive identification of Bartonella species.”
“We have studied the dynamic scanning of liquid-crystalline (LC) poly(p-phenyleneterephthalamide) sulfuric acid (PPTA-H(2)SO(4)) solution, and its blend with single-walled carbon nanotubes (SWNTs), by using a flat plate rotational rheometer. The effects of weight concentration and molecular weight of PPTA, as well as operating temperature, on dynamic viscoelasticity of the PPTA-H(2)SO(4) LC solution system are discussed. The transition from a biphasic system to a single-phase LC occurs in the weight concentration range of SWNTs from 0.1% to 0.2%, in which complex viscosity reaches the maximum at 0.2 wt% and the minimum MDV3100 order at 0.1 wt%, respectively, of SWNTs. With increasing SWNT weight concentration, the endothermic peak temperature increases from 73.6 to 79.9 degrees C. The PPTA/SWNT/H(2)SO(4) solution is in its plateau zone and storage modulus (G’) is a dominant factor within the frequency (omega) range of 0.1-10 rad/s. As omega increases, the G’ rises slightly, in direct proportion to the omega. The loss modulus (G ”) does not rise as a function of omega when omega < 1 s(-1), then when omega > 1 s(-1) G ” increases faster than
G’, yet not in any proportion to the omega.”
“Background: Dengue fever is the most important vector-borne viral disease. Four serotypes of dengue virus, DENV1 to DENV4, coexist. Infection by one serotype elicits long-lasting immunity to that serotype but not the other three. Subsequent infection by a different serotype is a risk factor for severe dengue. Domain III (ED3) of the viral envelope protein interacts with
cell receptors and contains epitopes recognized by neutralizing antibodies. We determined the serotype specificity and cross-reactivity of human IgMs directed against ED3 by using a well-characterized collection of 90 DENV-infected and 89 DENV-uninfected human serums.\n\nMethods: The recognitions between the four serotypes of ED3 and the serums were assayed with an IgM antibody-capture ELISA (MAC-ELISA) and artificial homodimeric antigens. The results were analyzed with selleck compound Receiving Operator Characteristic (ROC) curves.\n\nResults: The DENV-infected serums contained IgMs that reacted with one or several ED3 serotypes. The discrimination by ED3 between serums infected by the homotypic DENV and uninfected serums varied with the serotype in the decreasing order DENV1 > DENV2 > DENV3 > DENV4. The ED3 domain of DENV1 gave the highest discrimination between DENV-infected and DENV-uninfected serums, whatever the infecting serotype, and thus behaved like a universal ED3 domain for the detection of IgMs against DENV.
Conclusion: Taken together our data supports a strongly positive association between mitochondrial complex I gene variations and MS pathogenesis in a Filipino population. (C) 2014 Elsevier B.V. All rights reserved.”
“The development of automated, high-throughput organic synthesis and screening techniques has created an urgent demand for methods that rapidly determine the enantiomeric composition of chiral compounds. Enantioselective fluorescent sensors offer the potential for real-time, high-sensitivity techniques for determining enantiomeric data in high-throughput chiral assays. In this Account, we describe a range of fluorescent sensors derived from 1,1′-bi-2-naphthol (BINOL), Selleck TPCA-1 a readily
available biaryl compound with axial chirality.\n\nWe show that BINOL can be used to construct structurally diverse, chiral fluorescent sensors to carry out highly enantioselective, sensitive recognition of chiral WH-4-023 molecular weight amino alcohols, alpha-hydroxycarboxylic adds, and amino add derivatives. For example, we prepared an (S)-BINOL derivative whose 3,3′-positions are attached to two chiral amino alcohol units, each having two phenyl substituents. This compound shows a fluorescence enhancement of 950-fold in the presence of (S)-mandelic add but
very little change in the presence of (R)-mandelic acid. It also allows the enantiomers of this alpha-hydroxycarboxylic add to be visually discriminated by an enantioselective precipitation process.\n\nA structurally similar (S)-BINOL-amino alcohol molecule, but with three rather than two phenyl substitutents in each of the two amino alcohol units, was found to exhibit generally enantioselective fluorescence responses toward structurally diverse alpha-hydroxycarboxylic adds. We further
prepared a pseudoenantiomeric analogue of this compound from (R)-H8BINOL, selleck inhibitor which has the opposite chiral configuration at both the biaryl center as well as the pendant amino alcohols. These two compounds have opposite enantioselectivity in the recognition of a chiral substrate, with distinctly different fluorescence emission wavelengths. By mixing them together, we developed a pseudoenantiomeric sensor pair to facilitate chiral assays. Using this pseudoenantiomeric sensor pair allows both the concentration and the enantiomeric composition of a substrate to be determined in a single fluorescence measurement.\n\nWe synthesized another compound by ligating a terpyridine unit to BINOL and found that coordination of a Cu(II) ion to the terpyridine unit almost completely quenched its fluorescence. Displacement of the Cu2+ ion from this complex by chiral amino alcohols leads to enantioselective fluorescence enhancement. This BINOL-terpyridine-Cu(II) complex also exhibits enantioselective gel collapsing in the presence of chiral amino alcohols, providing a new visual chiral discrimination method.