P2Y12 mRNA transcripts were detected [25],

but receptor expression was not detected in the anterior pituitary cells (Yu et al., 2011). Currently most compounds to which patients will be exposed for more than 6 months duration must be evaluated for carcinogenicity potential during drug development (ICHS1A:). The two-year rat carcinogenicity bioassay, as outlined in the International Conference on Harmonization guidance documents (ICH S1, S2, S3), is used in conjunction with other assays to determine the carcinogenicity potential BTK pathway inhibitor of compounds. Human patient safety risk (if any) is determined based on the human relevance framework [6], [11] and [32]. This framework leverages two concepts to determine a statement

of confidence regarding patient safety risk: 1) is the weight of evidence sufficient to establish the mode of action (MOA) in animals and 2) is the MOA plausible in humans. Therefore, determining the MOA of a carcinogenicity finding is critical to accurately determine GSK269962 datasheet the human relevance of any findings from the carcinogenicity bioassays. The human relevance framework helps classify the human patient safety risk from high confidence in the rodent carcinogenicity data translating into patient safety risk, to the mechanism of action studies determining the rat carcinogenicity data has a MOA not plausible in human and thereby no patient safety risk. For example, central-acting dopamine agonists altered tumor incidences in rats is an example of lack of confidence in the MOA translating into human (Figure 1). This is because altered brain dopamine levels inhibit pituitary prolactin release in both female rats and humans but the decreased prolactin level alters tumor incidences of reproductive organs in female rats and not in humans as prolactin is luteotrophic in rats, but not in primates (Neuman, 1997). [6] termed this lack of confidence as being due to qualitative species differences. Therefore, the objectives of these studies were to (1) evaluate the Ticagrelor rat two-year carcinogenicity bioassay data, (2) investigate potential mode of action

PLEK2 (MOA) for any altered tumor findings and (3) interpret the data using the human relevance framework to determine the patient safety risk. All procedures were approved by the appropriate institutional Animal Care and Use Committee (IACUC) in accordance with The Guide for the Care and Use of Laboratory Animals. Rats were housed, as outlined within each experiment, with food and water provided ad libitum, unless otherwise stated. A standard light-dark cycle was maintained with a timer-regulated light period from 0600 to 1800 hours. The procedures within this study were consistent with the guidelines of the EU, US FDA and Japanese MHLW; prospective FDA protocol concurrence was sought and received under the Special Protocols procedure (ICHS1A).

The fact that the dermal LD50 of fipronil is higher than 2000 mg/kg bw [46] agrees with this observation. This kinetic profile might help to explain the three hours onset of behavioral effects observed in our pilot studies. As opposed to fipronil, others pesticides act in mammals in their original molecular form

and have their effects diminished after metabolism. Thus, future research is important to study the implications of kinetic parameters this website on risk assessment for neurotoxicity by these compounds. In conclusion, since non-target organisms are evidently exposed to the insecticides because of colocalization, it is important to have more information about their undeliverable effects. The present study confirmed that the insecticide fipronil has central behavioral effects in rats. Further studies with pirazole insecticides, including fipronil, are necessary to verify their neurotoxic potential in humans because of accidental and professional selleck exposure. “
“Drug-induced toxicity is a serious problem affecting patients undergoing chemotherapy. Depending on the toxicity profiles of individual drugs, therapeutic index may be limited, resulting in higher rate of treatment failures [1]. Apart from toxicity, cancer cells

also acquire self-remedial escape mechanisms such as drug efflux pumps or increased drug metabolisms devouring attack from chemotherapy, resulting in selleck kinase inhibitor the chemoresistance [2]. Doxorubicin (Dox) is a common chemotherapeutic drug with wide spectrum of anticancer activity against several malignancies. But, the most common side-effects associated with

anthracycline analogues like Dox include acute and chronic toxicities such as myelosuppression, cardiomyopathies and congestive cardiac failure [3] and [4]. To overcome these side-effects, integrated approaches utilizing combination therapies with cytotoxic, chemosensitizing and nanoparticle agents have been devised. Encapsulation of Dox in the form of PEGylated liposomes (Doxil) and Abraxane have increased the intratumoral delivery without much toxicity [3] and [5]. Dox conjugation with hydrophilic polymers was found to increase cytotoxicity by ‘enhanced permeation and retention’ (EPR) relative to free doxorubicin [6]. EPR effect enabled polymeric-drug nanoparticles to enhance their diffusion rate, and thus accumulate within tumor tissues than normal tissues, leading to enhanced antitumor efficacy and reduced side-effects. A small number of advanced drug delivery systems for Dox have been approved by the FDA for the treatment of ovarian cancer and Kaposi’s sarcoma which are in clinical use in the United States [7]. Still, there are substantial challenges like high treatment failure rates, unpredictable disease outcome, and tumor recurrence apart from toxicity, while using any single-agent drugs.

3, p = 0.01 at voxel level, and a cluster size probability of p < 0.05. Identifying sensorimotor activation in response to printed words often requires the increased power CX-5461 chemical structure of region of interest (ROI) analyses ( Willems & Casasanto, 2011). Therefore, two complementary ROI analyses

were performed in addition to a whole brain analysis. In a first set of ROI analyses, group average ROIs were derived from significant tool or animal category-specific clusters within each age group’s average activation map. For each individual within the group, mean BOLD responses to tool and animal words and pictures were then extracted from these group-specific ROIs. The advantage of this selection procedure is that it allows for straightforward identification of age-appropriate ROIs. A limitations of this approach, however, is that category selective responses underlying mean activations may be more variable at younger ages, so average

activation clusters may be less representative of individual activation patterns in earlier childhood (Poldrack, 2010). In addition, due to thresholding, different combinations of tool- and animal selective areas are grouped into single ROI clusters in different age groups, rendering comparisons across age for a given tool or animal region difficult to interpret. To account for these factors, an additional set of ROIs was defined consisting of category-selective voxels in pre-defined cortical regions within the individual activation maps. To select cortical PR-171 concentration areas with category-selective voxels in each individual activation map, we first created eight large spherical volumes (15 mm diameter) centred on average peak voxels or centre

of gravity coordinates of tool- or animal selective areas reported in click here the literature. The spheres were located in the tool picture selective left AIP (x = −44, y = −37, z = 44), left IFG (x = −46, y = 13, z = 14) left LOC/MTG (x = −48, y = −60, z = −4.1) ( Valyear, Cavina-Pratesi, Stiglick, & Culham, 2007) and the left and right medial FFG x = −25, y = −57, z = −7 and x = 22, y = −57, z = −5 ( Chao et al., 1999 and Devlin et al., 2005), and in the animal picture selective left and right lateral FFG: x = −38, y = −58, z = −12 and x = 36, y = −58, z = −12 ( Chao et al., 1999 and Devlin et al., 2005) and right posterior LOC, x = 46, y = −70, z = −1 ( Grill-Spector, Knouf, & Kanwisher, 2004; Peelen & Downing, 2005). Crucially, previous findings ( Dekker et al., 2011) corroborated by the current results, suggest that the overall organisation of tool and animal-selective areas across the brain is qualitatively adult-like by 6 years of age, and hence that identifying tool and animal picture-selective voxels of adults and children in the same cortical regions, is appropriate in this case.

Mit den heute www.selleckchem.com/products/17-AAG(Geldanamycin).html verfügbaren modernen Methoden ist es möglich und unerlässlich, genetische und epigenetische Studien einzubeziehen, um die individuellen Manifestationen des Manganismus besser zu verstehen, die von den unterschiedlichen Bedingungen der Mn-Exposition sowie von Geschlecht, Alter und Umwelt abhängig sind. Eine Literaturübersicht zur

Mn-Speziation im Hinblick auf Neurodegeneration und in Übereinstimmung mit den IUPAC-Definitionen der Speziation, die von Templeton et al. [90] publiziert wurden, ergab, dass die Mn-Speziation mit Blick auf neurodegenerative Effekte ab dem Jahr 2004 [91] hauptsächlich von unserer Gruppe durchgeführt wurde, was zu einer Reihe aufeinanderfolgender Publikationen führte, von denen die ersten im Jahr 2007 zusammengefasst wurden [9]. Das wichtigste Ergebnis dieser Arbeiten war, dass in menschlichem Serum vor allem Mn-Verbindungen mit hohem Molekulargewicht (HMM) vorkamen, die der learn more α-2-Makroglobulin- und der Transferrin-/Albumin-Fraktion zuzurechnen sind, und nur wenige Mn-Spezies mit

niedrigem Molekulargewicht (LMM), während im Liquor hauptsächlich LMM gefunden wurden, wobei Mn-Citrat gegenüber einigen anderen überwog. Folglich wurde die Hypothese formuliert, dass Mn-Citrat nach einer Mn-Exposition eine äußerst wichtige Mn-Spezies darstellen könnte, die die neuronalen Barrieren ohne ausreichende Kontrolle passieren kann [9] and [92]. Seit 2007 wurden in verschiedenen Folgestudien zur Mn-Speziation die noch offenen Fragen im Zusammenhang mit Mn-Spezies untersucht. Folgende Fragen wurden untersucht: (a) Wie hoch sind die Konzentrationen von Mn-Spezies an den neuronalen Barrieren, d. h. direkt davor (im Serum) und dahinter (im Liquor)? Nischwitz et al. [57] befassten sich mit

den Fragen (a) und (b): Diese Autoren untersuchten die Permeabilität der Blut-Liquor-Schranke für ausgewählte Metalle (Mn, Fe, Cu, Zn, Mg und Ca). Während der Speziationsanalyse war es ein Problem, die Stabilität der Mn-Spezies aufrechtzuerhalten. Daher wurde durchgehend die Methode der Größenausschlusschromatographie in Kombination mit Massenspektrometrie mit induktiv gekoppeltem Plasma (ICP-MS) verwendet. Peakfraktionen in Serum und Liquor wurden quantifiziert, PDK4 und die Liquor/Serum-Quotienten wurden berechnet. Das wichtigste Ergebnis dieser Studie war, dass hinsichtlich der molekularen Größenverteilung der Spezies der ausgewählten Metalle signifikante Unterschiede zwischen den Liquor- und den Serumproben auftraten. Es wurde angenommen, dass dies auf die selektive Permeabilität der BCB für Metallspezies aus dem Serum in den Liquor zurückzuführen war. Was Mn betraf, so war der Gradient vom Serum zum Liquor für alle Spezies negativ, außer für die Mn-Citrat-Fraktion, die signifikant angereichert war. Im Serum waren Fe, Cu und Zn hauptsächlich an HMM-Spezies gebunden, Mg und Ca dagegen an LMM-Spezies.

For each passage, in average fifteen to twenty cells were analysed. For detection of surface antigen, adherent cells were detached with 0.25% trypsin solution (Invitrogen), washed with saline and incubated at 4 °C for 30 min with following antibodies diluted 1:100: biotin anti-mouse CD31 (BD Biosciences Pharmingen, San Diego, CA, USA), biotin anti-human stromal stem cells – STRO-1 (R&D Systems, Minneapolis, MN, USA), PE anti-mouse CD34 (Invitrogen), PE anti-mouse/human oct-4 (BD Pharmingen), PE anti-mouse CD73 (BD Pharmingen), PE anti-mouse CD90 (Invitrogen), PE anti-mouse CD11b (BD Pharmingen), PE anti-mouse CD44 (BD Pharmingen), PE anti-mouse CD117 (Invitrogen), APC anti-mouse CD45 (Invitrogen),

selleckchem PE-Cy5.5 anti-mouse stem cell antigen – Sca-1 (Invitrogen) or 0.5 μg/mL propidium iodide (BD Pharmingen). Excess antibody was removed by washing. Streptavidin PE-Cy5.5 diluted 1:100 (BD Pharmingen) was used after biotin antibody. Cells were fixed with 1% formaldehyde. Quantitative find more evaluation of the exponential cell expansion was estimated by Carboxyfluorescein succinimidyl ester – CFSE assays (Invitrogen/Molecular Probes). CFSE staining was performed according to methodology previously described.16 The acquisition and analysis were done using a FACScalibur cytometer

(Becton Dickinson, San Diego, CA, USA) with the CellQuest software. At least 50,000 events were collected. Alkaline phosphatase expression was evaluated in monolayers of cells in the third passage cultivated in 24 well plates. USP-1, a mouse embryonic stem cell line17 was used as a positive control. Cultures were Carbohydrate washed with PBS, fixed with 4% paraformaldehyde (Sigma) in PBS, washed with rinse buffer, and stained with a mix fast red violet (FRV) with naphthol phosphate solution and water as described in the protocol of the embryonic stem cell characterization kit (Millipore Corporation, Billerica, MA). Positive alkaline phosphatase expression was identified by red cell colonies visualized using an inverted optic microscope (Olympus). For immunofluorescence analysis, 13-mm diameter glass coverslips (Knittel, Braunschweig, Germany)

were placed in a 24-well plate and mDPSC (5 × 106) were added in each well. Cells were washed in PBS 1×, fixed with 4% paraformaldehyde and permeabilized with 0.1% triton X-100 for 10 min. After blocking with PBS containing 5% BSA (Sigma), the cells were incubated with primary antibodies diluted 1:100. The embryonic stem cell characterization kit (Chemicon, Temecula, CA, USA) was used for detection of the following primary antibodies: SSEA-1 (stage-specific embryonic antigen-1; IgM monoclononal antibody), SSEA-4 (IgG monoclononal antibody), TRA-1-60 (keratin sulfate-associated antigens; IgM monoclononal antibody). After washing, appropriate secondary antibodies goat anti-mouse IgG or IgM Alexa Fluor 568 (Invitrogen/Molecular Probes) diluted 1:200 were added in the well.

In cells, enzymes often exist in multiple forms arising from the same (splice variants) or different loci in the genome. In contrast, in a reconstituted UK-371804 price biochemical system, the enzyme is often isolated, lacking many or all of its native binding partners, which can significantly affect enzyme stability and activity in vitro. Additionally, it may be difficult to express and purify the enzyme in its native form due to size and/or

stability limitations in the absence of these partner proteins. Hence, numerous constructs are often attempted in the expression and purification of the desired enzyme, including truncated variants and alternatively tagged species in an attempt to maximize protein yield, stability and activity. A caveat of these artificial alterations is that the more one diverges from the natural protein, the more likely it is to identify

compounds that lack a physiologically relevant mechanism and to miss compounds that work under physiological conditions. The choice of which protein construct to employ for development of the enzyme assay depends on several factors. Initial tests that assess differences in both the activity and stability of protein constructs are critical in deciding which constructs to advance. In addition, the use of “tool” compounds, that is compounds with known modes of inhibition (MoI), can be extremely revealing in evaluating which construct to PtdIns(3,4)P2 ultimately use in a HTS selleck chemicals based on the desired MoI. When multiple constructs of an enzyme use the same substrate, it is possible to compare their activities using the Michaelis–Menten

constants in the form of kcat/Km. This takes into account both the rate determining step which limits the maximal velocity of the enzyme reaction (expressed in the constant kcat) and the propensity of the substrate to be turned over to product (Km). While subtle differences in the rate and or Km may exist among constructs, large differences in kcat/Km can indicate significant differences in the structure and/or stability of the construct. Additionally, the specific activity can also be used to compare different preparations of the same enzyme construct. The specific activity is the Vmax (calculated at saturating amounts of substrate) divided by the mass of enzyme in the reaction and is usually expressed in μmol min−1 mg−1. A severe decrease in assay performance may be indicative of poor batch reproducibility or indicate a decay in batch activity which can be checked by monitoring the specific activity between different enzyme batches or different samples of enzymes from the same batch. The purity of the enzyme target must also be considered, as the use of an impure enzyme preparation can lead to the selection of aberrant or mis-targeted inhibitor compounds. Enzyme purity can be assessed in a number of ways.

, 2005, Zhang et al., 2008, Milanski et al., 2009, Posey et al., 2009 and Thaler et al., 2012). However, whether obesity-associated central inflammation per se contributes to HPA axis dysfunction is unclear, as no study has

yet dissected hypothalamic inflammation in the context of obesity to this degree of detail. It is worth noting, central inflammation in contexts other than obesity certainly leads to HPA axis dysfunction. For instance, brain pro-inflammatory cytokine levels and other inflammatory markers are elevated in rodent models of depression ( Patki et al., 2013). Chronic stress can lead to increased hypothalamic pro-inflammatory cytokine (IL-1β, TNFα) expression, as well as to the recruitment of peripherally-derived monocytes click here (bone marrow-derived immune cells that will differentiate into macrophages upon entry into tissues) into this website the brain, including to anxiety-related brain regions such as the amygdala ( Johnson et al., 2002 and Wohleb et al., 2013b). This effect is linked to anxiety-related behavior and potentially to anxiety-related mood disorders, with a stress-sensitized monocyte response contributing to excessive anxiety in mice previously exposed to chronic social defeat (

Wohleb et al., 2013a and Wohleb et al., 2013b). ICV LPS leads to PVN activation and an increase in IL-1β in this region and an increase in arginine vasopressin ( Xia and Krukoff, 2003). In addition, icv IL-1β administration induces muscle atrophy in mice and this effect is mediated by the HPA axis ( Braun et al., 2011). Notably, the PVN and ARC are also reciprocally interconnected, as are feeding and stress ( Shin et al., 2009). Thus, any adverse effect on the ARC, including inflammation, can potentially Carnitine palmitoyltransferase II affect PVN function even if inflammation does not occur in the PVN itself. It is now apparent that our sensing and regulation of food intake is not simply determined by the ARC sensing peripheral

nutritional signals and translating these into an ‘eat/do not eat’ command. Rather, the ARC is intimately connected with other regions of the hypothalamus and the rest of the brain to integrate the body’s nutritional needs with the external environment and the body’s other demands (Shin et al., 2009). Thus, the hypothalamus is closely connected with motivation and reward pathways in orbitofrontal cortex, hippocampus, mesolimbic dopamine system, nucleus accumbens, striatum and prefrontal cortex, as well as sensory and memory systems (Shin et al., 2009). Thus, inflammation in the ARC that disrupts feeding signals there will undoubtedly also disrupt signaling to, and potentially from, these brain regions and thus potentially impair cognitive function (Fig. 1). There is currently a good body of evidence to suggest high fat feeding leads to inflammation within the hypothalamus. However, fewer studies have examined if this inflammation is specific to hypothalamus or extends into other brain regions.

Less frequent stimulation of the network did not have any qualitative effect on its dynamics. In the second approach, the incorporation AZD1208 molecular weight of augmentation, i.e. slow synaptic facilitation (Wang et

al., 2006, see Experimental procedures) added new functional aspects to the dynamics. These simulations correspond to memory processes involved in a memory replay phenomenon, which can be linked to multi-item working memory maintenance in the cortex (Fuentemilla et al., 2010). A specific memory pattern was stimulated at first, as in the previous simulation paradigm, and after the resulting brief initial activation the internal dynamics of the network caused this particular attractor state to periodically reactivate without any successive external stimulation (Mongillo et al., 2008, Lundqvist et al., 2011 and Lundqvist et al., 2012). This occurred since the synaptic augmentation had a longer time constant than the synaptic depression. During periods of activity in the recurrent network these two synaptic mechanisms balanced each other out, but once the memory retrieval terminated synaptic depression started decaying faster. As a result, synaptic conductances of the excitatory recurrent connections of the recently terminated patterns became temporarily boosted. This way 5.0±0.7 (mean±standard deviation, 100 trials) memory items could be encoded by initial NVP-BKM120 mouse sequential activation of the corresponding attractor states

followed by spontaneous periodic reactivations

of these specific patterns. Fig. 2B illustrates a spike raster obtained during part of a trial with periodic reactivations. Trials were typically run for 20 s to obtain reliable statistics. We analyzed both the spiking activity and the synthesized LFP signals collected during simulated memory processes implicated in memory pattern completion or sequential memory replay phenomena. During periods of interleaved idling in the non-coding ground state and memory activation we found in both types of memory simulations distinct frequency components in the power spectrum corresponding to the upper alpha/lower beta oscillations and the coupled (Chrobak and Buzsaki, 1998; Palva et al., 2005 and Canolty et al., 2006) theta- (2–5 Hz) and gamma- (25–35 Hz) band activity (Fig. 2C and D). The nested theta and gamma oscillations accompanied the coding attractor states (Fig. 3). In cued trials, an additional ~10 Hz MycoClean Mycoplasma Removal Kit alpha rhythm coupled to the gamma and theta emerged in the synthesized LFPs (Fig. 2C). Finally, the aforementioned upper-alpha-/lower-beta-band activity (15−20 Hz) was manifested as an attribute of the idling non-coding ground state. Here however we only focused on the nested oscillations during memory retrieval in the coding attractor states. The coherence analysis performed on the LFPs within as well as between hypercolumns revealed a generally decreasing trend with both distance and frequency. Spiking, although highly irregular for single cells (Lundqvist et al.

Kind regards, Ursula Culligan “
“Over the last decades, the use of polymers as food packaging materials has increased considerably due to their advantages over other traditional materials such as glass or

tinplate. A selleck screening library great advantage of plastics is the large variety of materials and compositions available, so the most convenient packaging design can be adapted to the very specific needs of each product (López-Rubio et al., 2004). However, it is widely accepted that the use of long-lasting polymers for short-lived applications (packaging, catering, surgery, hygiene) is not entirely adequate (Avérous, 2004). A huge amount of garbage is generated daily, in which food packaging check details plays a considerable part. This waste is composed of many different types of material, some of which is not biodegradable and will remain without decomposing for hundreds, sometimes thousands of years. In this context, the development of biodegradable films (BF) for packaging materials that can be used as a substitute for petrochemical polymers is an interesting perspective, since it provides an alternative to non-degradable products, and increases income in the agricultural sector (Souza, Ditchfield, & Tadini,

2010). Starch has been considered as one of the most promising candidates for the future primarily because of an attractive combination of its large availability and relatively low price (Chivrac, Angellier-Coussy, Guillard, Pollet, & Avérous, 2010). Cassava starch has been extensively used to produce BF (Alves et al., 2007, Chen and Lai, 2008, Chillo et al., 2008, Famá et al., 2006, Famá et al., 2007, Flores et al., 2007, Henrique et al., 2008, Kechichian et al., 2010, Mali et al., 2006, Müller et al., 2008, Paes et al., 2008, Parra et al., 2004, Shimazu et al., 2007, Teixeira et al., 2007, The OSBPL9 et al., 2009,

Veiga-Santos, Oliveira et al., 2005, Veiga-Santos, Suzuki et al., 2005 and Veiga-Santos et al., 2008) and the results indicated that these carbohydrates are promising materials in this regard (Müller et al., 2008). Films developed from starch are described as isotropic, odorless, tasteless, colorless, non-toxic and biologically degradable (Flores et al., 2007). Unfortunately, there are some strong limitations for developing starch based products, since they present poor tensile properties and high water vapor permeability when compared to conventional films derived from crude oil (Souza et al., 2010) on account of their hydrophilic nature and their sensitivity to moisture content, a factor that is difficult to control (Wilhelm, Sierakowski, Souza, & Wypych, 2003).

The AW approach holds for slower motional rates k=3kHz, but the agreement becomes worse at higher rates. Another example is shown Fig. 4c, which features the same

comparison for the case of a CH3CH3 group executing two-site jumps with reorientation angle of 109°109°, including an internal fast permutation of the CH3CH3 protons. This corresponds to the motion executed by the CH3CH3 groups in dimethyl sulfone (DMS) molecular crystals, of course assuming the protons permutation belonging to the methyl group to be in the fast limit. Again, the AW approximation is not suitable to describe the curve for rates higher than 3kHz. Cases of molecular motions with different geometries and numbers of sites were tested and similar results were found. To understand the reason why the AW approximation is adequate for describing Pirfenidone concentration the 2tr-tC-recDIPSHIFT2tr-tC-recDIPSHIFT curves of the CH groups, but fails in the case of CH2CH2 and CH3CH3, in Fig. 5 and Fig. 6 we address the fidelity this website of the Gaussian approximations (dashed

blue lines) for reproducing the general features of the local dipolar field distribution (solid black lines) for CH and CH2 groups, respectively. The corresponding dipolar spectra were in each case calculated for the (a) rigid and (b) fast motion limits, considering the motion geometries displayed as inset in Fig. 4. In the rigid limit, both CH and CH2CH2 dipolar powder patterns, Fig. 5 and Fig. 6, resemble unimodal

distributions, so a single second moment can be used in Eq. (4). However, as demonstrated in Fig. 5 and Fig. 6, in the fast-motion limit the pattern for the CH group is still well represented by a single Gaussian, but the pattern for the CH2CH2 group is clearly composed of two components, i.e., a Pake pattern of about 20 kHz width and an isotropic line. The former arises from the two parallel spin configurations of the two Dimethyl sulfoxide protons, while the latter arises from the antiparallel configurations [48], for which the coupling cancels for the given case of identical dipolar tensors arising from the motional averaging. Thus, the δ  -function shaped “central transition” in this spectrum has the same integral intensity as the broad Pake pattern. A similar behavior regarding the bimodal spectrum is also observed for the case of CH3CH3 groups. As the core of the AW approximation is that the given frequency distribution can be modeled as a Gaussian, it is straightforward to rationalize the observed behavior, where the description is accurate in describing the 2tr-tC-recDIPSHIFT2tr-tC-recDIPSHIFT data of CH groups, but fails for the case of CH2. This suggests that the scenarios for which the AW approximation is not completely satisfactory (CH2 and CH3) may be improved by increasing the number of Gaussian functions used to describe the local field, as demonstrated by the red dotted lines in Fig.