1 in kinetic resolution of 4-nitrophenyl 2-methylheptanoate Enzy

1 in kinetic resolution of 4-nitrophenyl 2-methylheptanoate. Enzyme variants were expressed in Pichia pastoris by using the episomal vector pBGP1 which allowed efficient secretory expression check details of the lipase. Iterative rounds of CASTing yielded variants with good selectivity toward both the (S)- and the (R)-enantiomer. The best obtained enzyme variants had E-values of 52 (S)

and 27 (R).”
“1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) causes nigrostriatal dopaminergic neurotoxicity and behavioral impairment in rodents similar to Parkinson’s disease. The MPTP mouse model is widely used to evaluate new protective agents. EGb 761 is a well-defined mixture of active compounds extracted from Ginkgo biloba leaves according to a standardized procedure. We have shown that EGb 761 attenuates the loss of striatel dopamine levels and prevents the neurodegeneration of the nigrostriatal

pathway induced by MPTP. This finding shows that neuroprotective effects of EGb 761 act, in part, on the dopamine system. Therefore, this study investigates whether EGb 761 exerts dopaminergic neuroprotection through the regulation of dopamine-related gene expression in MPTP-induced Parkinsonism. find more Male C57BL/6J mice were injected with MPTP (30 mg/kg, i.p.) for 5 days and later with EGb 761 (40 mg/kg, i.p.) daily for 18 days. The expression of selected genes was evaluated in the striatum and midbrain by quantitative PCR. The genes for tyrosine hydroxylase (Th), vesicular monoamine

transporter 2 (Vmat2), dopamine transporter (Dat), dopamine D2 receptor (Da-d2r), and transcription factors (Pitx3 and Nurr1) related to dopamine neurotransmission were selected for the analysis. EGb 761 administration to MPTP-treated mice protected Th (41%), Vmat2 (15%), Dat (102%), Da-d2r (46%), Pitx3 (63%), and Nurr1 (148%) mRNA levels in the midbrain, all of which were up-regulated. However, EGb 761 partially reversed the MPTP effect exclusively for Th (48%) and Nurrl (96%) mRNA in the striatum. Only Th and Nurr1 mRNA and protein levels were regulated”
“Although an important role for mast cells in several Flavopiridol (Alvocidib) viral infections has been demonstrated, its role in the invasion of highly pathogenic H5N1 influenza virus is unknown. In the present study, we demonstrate that mast cells were activated significantly by H5N1 virus (A/chicken/Henan/1/2004) infection both in vivo and in vitro. Mast cells could possibly intensify the lung injury that results from H5N1 infection by releasing proinflammatory mediators, including histamine, tryptase, and gamma interferon (IFN-gamma). Lung lesions and apoptosis induced by H5N1 infection were reduced dramatically by treatment with ketotifen, which is a mast cell degranulation inhibitor. A combination of ketotifen and the neuraminidase inhibitor oseltamivir protected 100% of the mice from death postinfection.

The successful detection of MCMV in maize seeds suggested the fea

The successful detection of MCMV in maize seeds suggested the feasibility of this procedure for routine testing. (C) 2010 Elsevier B.V. All rights reserved.”
“You are queuing at the bus stop, and notice that someone suddenly turns her walk into a run: typically, you assume that she wants to catch the bus and may want to tell the driver to wait. Faced with a sudden speed change, rather than considering it bizarre or unnatural, observers attach a meaning to it, and act consequently. In a social context, speed of a movement often bears as much significance as its form,

and can be adapted to vehicle Sapanisertib in vitro precise meanings. This pragmatic rule facilitates decoding of non-verbal messages from other individuals, but may not necessarily apply when observing one’s own movements, for which intentions should be informative enough. Hence, the range of motion speeds labeled as ‘natural’ could be broader for other people’s actions compared to one’s

own. We explored this possibility through a task in which human observers decided whether speed of a gesture had been artificially modified. A virtual hand was presented, which – unbeknownst to participants – moved according to the kinematics of either the observer, or another individual. Although a self/other distinction was never required, participants applied different criteria when dealing with self compared to other people’s gestures, suggesting that the brain implicitly extracts identity information before any overt judgment is produced. Interestingly, observers PD173074 order were reluctant to labeling movements of another individual as artificial, in keeping with the hypothesis that large variations in movements’ speed can vehicle social messages, and therefore are not regarded a priori as unnatural. (C) 2011 Elsevier Branched chain aminotransferase Ireland Ltd. All rights reserved.”
“Recombinant poxviruses are important tools for research and some are candidate vaccines. To make these viruses a simple, small vector that can be used to engineer

multiple strains of vaccinia virus and other model poxviruses, including ectromelia virus is of value. Here a set of plasmids and methods for making these viruses that uses an enhanced green fluorescent protein-blasticidin resistance (GFP-bsd) fusion gene as a transient selectable marker are described. This gene is smaller than any of the bi-functional selection markers used previously. The versatility of the method across different poxviruses is demonstrated by engineering changes into multiple loci of the WR and Modified Vaccinia Ankara (MVA) strains of vaccinia virus and also ectromelia virus. Finally, a set of vaccinia virus sequences for directing homologous recombination that are very highly conserved was designed and tested. These sequences allow a single plasmid to be used to insert a transgene into multiple strains of the virus. (C) 2010 Elsevier B.V. All rights reserved.


“Objectives: Extensive aneurysms of the ascending, arch, a


“Objectives: Extensive aneurysms of the ascending, arch, and descending thoracic aorta traditionally have been managed with a 2-stage “”elephant trunk” procedure. Single-stage transmediastinal

repairs have also been used, because in some patients the entire repair is not completed owing to either complications during first-stage repair or magnitude of the second-stage operation. Since 2006, second-stage elephant trunks have been preferentially completed with endovascular stent grafts in anatomically appropriate patients. This study compares outcomes of 2-stage elephant trunk, single-stage, and hybrid endovascular repairs of extensive thoracic aortic aneurysms.

Methods: This is a single-institution retrospective cohort study of 103 patients (1992-2007) with extensive thoracic aortic aneurysms undergoing 2-stage elephant selleck products trunk repair with either surgical (OPEN) or endovascular (TEVAR) completion versus single-stage transmediastinal replacement (SS). Outcomes were analyzed with Statistica 8.0 software

(Tulsa, Okla).

Results: Of 103 patients, 74 had elephant trunk procedures, 24 TEVAR-eligible and 50 OPEN, and 29 had SS. Completion rates were higher with TEVAR than OPEN (78% vs 47%; P = .01). Seven of 18 TEVARs were performed during the index hospitalization. TEVAR patients had shorter second-stage hospital stay (5.5 vs 16.5 days [P < .01]), required fewer transfusions (P < .01), and had less acute kidney injury (P= .04). There were no differences in mortality, paraplegia, or stroke. Six-month Kaplan-Meier survival estimates for OPEN, TEVAR, and SS were 64%, 78%, and 64% find more (P = .08).

Conclusion: More patients complete the second stage when TEVAR is used after elephant trunk repair, with fewer hospital days and transfusions. TEVAR is feasible and safe in the hybrid management of extensive thoracic aortic aneurysms.”
“Objective: PD184352 (CI-1040) The optimal management of moderate functional mitral regurgitation at the time of aortic valve replacement remains undefined.

Methods: We retrospectively

identified 686 consecutive patients undergoing aortic valve replacement between 1993 and 2006 with at least moderate (grade 2 or more) functional mitral regurgitation. Patients with structural valve abnormalities or significant coronary artery disease were excluded, leaving 190 in the study. Analyses for predictors of residual mitral regurgitation and survival were performed. The impact of mitral regurgitation on survival was further analyzed among 91 patients case matched for age, gender, and left ventricular ejection fraction to individuals without mitral regurgitation undergoing isolated aortic valve replacement.

Results: The mean age of the study group was 74 +/- 11 years, 45% were male, and 78% had New York Heart Association III or IV Class classification. The mean preoperative ejection fraction was 48% +/- 17%.

The striatal DA dynamics correlated with the CBV response These

The striatal DA dynamics correlated with the CBV response. These results demonstrate Oligomycin A that electrical stimulation near the median nerve activates Glu/GABA release, which subsequently attenuate excess striatal DA release. These data provide evidence for physiologic modulation caused by electroacupuncture at points near the median nerve. (c) 2007 Elsevier Ireland Ltd. All rights reserved.”
“We investigated the hypothesis that gemtuzumab ozogamicin (GO), an anti-CD33 immunotoxin would improve the efficacy of fludarabine/melphalan

as a preparative regimen for allogeneic hematopoietic stem cell transplantation (HSCT) in a phase I/II trial. Toxicity was defined as grades III-IV organ damage, engraftment failure or death within 30 days. ‘Response’ was engraftment and remission (CR) on day +30. We sought to determine the GO dose (2, 4 or 6 mg m(-2)) giving the best trade-off between toxicity and response. All patients were not candidates for myeloablative regimens. Treatment plan: GO (day – 12), fludarabine 30 mg m(-2) (days-5 to -2), melphalan 140 mg m(-2) (day – 2) and HSCT (day 0). GVHD prophylaxis was tacrolimus and mini-methotrexate. Diagnoses were AML (n = 47), MDS (n = 4) or CML (n = 1). Median age was 53 years (range, 13-72). All but PLX-4720 three patients were not in CR. Donors were related (n = 33) or unrelated (n = 19). Toxicity and response rates at

4 mg m(-2) were 50% (n = 4) and 50% (n = 4). GO dose was de-escalated to 2 mg m(-2): 18% had toxicity (n = 8) and 82% responded (n = 36). 100-day TRM was 15%; one patient had reversible hepatic VOD. Median follow-up was 37 months. Median event-free

and overall survival was 6 and 11 months. GO 2 mg m(-2) can be safely added to fludarabine/melphalan, and this regimen merits further evaluation.”
“The present event-related potential (ERP) study was conducted to investigate the P3 component in response to love-related stimuli while controlling for task-related factors, and to dissociate the influences of both love-related and task-related attention on the P3 amplitude. In an oddball paradigm, photographs of beloved and friends served as target and distractor stimuli. Love-related and task-related attention were separated by varying the target and distractor status of the beloved and selleck chemicals friends full factorially. As expected, the P3 amplitude was larger for beloved compared to friends and for targets compared to distractors. Moreover, task-related and love-related attention were unconfounded. These results are in line with findings that the P3 is modulated by both emotion- and task-related factors, supporting the view that the P3 amplitude reflects attention. Furthermore, this study validates the notion that romantic love is accompanied by increased attention for stimuli associated with the beloved, and also shows that this form of attention is different from task-related attention. (c) 2007 Elsevier Ireland Ltd. All rights reserved.

Here, we report that the CB(1) receptor antagonist rimonabant dis

Here, we report that the CB(1) receptor antagonist rimonabant disrupts extinction learning in an aversive, but not in an appetitive, Barnes maze conditioning task. Accordingly, these results provide CH5183284 chemical structure compelling support for the hypothesis that the endogenous cannabinoid system plays a necessary role in the extinction of aversively motivated behaviors but is expendable

for appetitively motivated behaviors.”
“Hemispheric lateralization is well known in the cerebral cortex, but not in subcortical structures like basal ganglia. The goal of our study was to determine whether lateralization was present in the direct and indirect striatal pathways. We studied gene expression in the striatum of healthy rats, which was divided into two sectors, medial and lateral. Dynorphin

(DYN) and enkephalin (ENK) mRNA were analyzed as markers of the learn more direct and indirect striatal pathways, respectively and glutamic acid decarboxylase (GAD) mRNA was analyzed as a marker of all medium spiny neurons. DYN and GAD mRNA expression was higher on the left hemisphere in the medial sector of the striatum, but not in the lateral one. We did not observe any difference between sides with ENK mRNA expression. We suggest the presence of a lateralization in the medial striatum, which is specific for the direct striatal pathway. (C) 2008 Elsevier Ireland Ltd. All rights reserved.”
“Spatial familiarization consists of a decrease in the exploratory activity over time after exposure to a place. Here, we show that a 30-min exposure to an open field led to a pronounced decrease in the exploratory behavior of rats, generating context familiarity. This

behavioral output is associated with a selective decrease in hippocampal PKM zeta levels. A short 5-min exposure did not induce spatial familiarity or a decrease in PKM zeta, while inactivation of hippocampal PKM zeta by the specific Nintedanib (BIBF 1120) inhibitor ZIP was sufficient to induce spatial familiarity, suggesting that the decrease in PKM zeta is involved in setting a given context as a familiar place.”
“The effect of general anesthetics on the developing brain is receiving growing attention. Nonetheless. there remains a paucity of evidence regarding the effect of sevoflurane, a widely used anesthetic in pediatric anesthesia. This study was designed to investigate the effect of sevoflurane on nerve cell apoptosis in the developing brain. Techniques to detect cell apoptosis included immunohistochemistry of cleaved caspase-3 and single-strand DNA, as well as electron microscopy. Elevated cleaved caspase-3 was also validated semi-quantitatively by immunoblotting assay. Mouse pups (day 7 postnatal) were subjected to sevoflurane inhalation. Twelve hours later, dramatically increased cleaved caspase-3 and single-strand DNA immunoreactivity were detected in the pup brains.

Despite clinical evidence for the efficacy of a beta-noradrenergi

Despite clinical evidence for the efficacy of a beta-noradrenergic receptor blockade with propranolol in the alleviation of anxiety symptoms and the secondary prevention of post traumatic stress disorder, preclinical evidence for a beta-noradrenergic modulation of BLA activity in humans is missing.

Method. We combined functional magnetic resonance imaging in healthy volunteers with probabilistic mapping of intra-amygdalar responses to fearful, neutral and happy facial expressions to test the hypothesis that a beta-noradrenergic receptor blockade ABT-263 purchase with propranolol would inactivate

the BLA.

Results. Consistent with our a priori hypothesis, propranolol diminished BLA responses to facial expressions, independent of their emotional valence.

The absence of activity changes in probabilistically defined visual control regions underscores the specific action of propranolol in the BLA.

Conclusions. Our findings provide the missing link between the anxiolytic potential of propranolol and the biological basis of beta-noradrenergic activation in the human BLA as a key target for the pharmacological inhibition of anxiety neurocircuitry. Moreover, our findings add to emerging evidence that NE modulates both the reactivity (sensitivity) and the operating characteristics (specificity) of the BLA via beta-noradrenergic receptors.”
“Previous EEG source localization studies in obsessive 3 Methyladenine compulsive disorder

(OCD) reported ambiguous results. The reason probably lies in different OCD samples included in the studies obsessive compulsive subjects selected based on a psychopathology questionnaire (the Symptom Checklist Revised), drug naive OCD cases or patients with a long-term disorder. This study was conceived as a replication of our previous research on OCD population coming to treatment in Prague Psychiatric Centre [9]. We included 50 OCD patients (8 drug-free and 42 medicated with SSRIs) and 50 healthy controls. All subjects were different from those enrolled in the previous study. Resting state EEG was analyzed in 8 frequency Cell press bands as well as with 1 Hz frequency resolution using the standardized low-resolution electromagnetic tomography (sLORETA). In OCD, sLORETA indicated low-frequency power excess at 2 and 3 Hz in the cingulate gyrus with maximal t-values in Brodmann area 24. The low-frequency activity was unrelated to the severity of clinical symptoms and illness duration but delta power in the right orbitofrontal cortex positively correlated with age of OCD onset. Our results confirm previous finding of the low-frequency excess in the cingulate gyrus in OCD and document the essential role of delta frequencies. Delta activity in the cingulate gyms is negatively associated with reward-signalling dopamine release in the ventral striatum and increases in states connected with a need for reinforcement.

The IL-18 labeled cells were largely colocalized with Iba1, sugge

The IL-18 labeled cells were largely colocalized with Iba1, suggesting this upregulation

occurred in hyperactive microglia. We also found that the IL-18 induction coexisted with phosphorylated p38 MAPK, indicating a possible role of p38 in the regulation of IL-18. Our findings are the first report that injury of trigeminal nerve induced IL-18 upregulation in activated microglia in the Vc, suggesting a possible role of IL-18 in orofacial neuropathic pain. (C) 2012 Elsevier Ireland Ltd. All rights reserved.”
“Human noroviruses (HuNoVs) are the major cause of epidemic, nonbacterial Berzosertib price gastroenteritis worldwide. Due to the lack of a tractable model system and the inability to grow HuNoVs in cell culture, factors required

for the norovirus (NoV) life cycle and pathogenesis in the host remain largely unknown. The discovery of murine norovirus (MNV) and the development of reverse-genetics systems for this virus provide an opportunity to study these aspects of NoV infection in vitro and in vivo. Previous studies identified a single amino acid at residue 296 in the protruding (P) domain of the capsid protein that is responsible for determining the virulence of the MNV clone MNV1.CW1 in 12956/SvEv background STAT1-deficient (STAT1(-/-)) mice. In this report, we identified and characterized another determinant of lethality in the P domain that is necessary and sufficient to determine the replication and pathogenesis of the MNV clones MNV1.CW3 and CR6.STL1 in C57BL/6 background STAT1(-/-) mice. Furthermore, we

describe how the 10058-F4 role of residue 296 in MNV virulence differs between STAT1(-/-) mouse strains. We also describe potential interactions between subdomains of the P domain, as well as between other virus elements, which facilitate recovery of MNV using a reverse-genetics system.”
“Increasing Urease data support an association of cyclothymic temperament with bipolarity, but our knowledge about the relationship of affective temperaments (ATs) to depressive symptoms based on inheritance in a non-clinical population is limited. The aim of this article was to derrionstrate how ATs and affective family history relate to the depressive symptoms in a general population. Subjects comprised 501 Hungarian adults who completed a background questionnaire, the TEMPS-A, the Zung Self-Rating Depression Scale (ZSDS) and the depression subscale of the Brief Symptom Inventory (BSI-D). Stepwise linear regression was performed to analyse the role of ATs and affective family history (AFH(0) and AFH(1)) in the variance of ZSDS and BSI-D scores. Cyclothymic, depressive and anxious temperaments have a significant role in the explained variance of depression scores, and they are all significantly related to AFH(1).

Productive infection of T cells by HIV is dependent upon the targ

Productive infection of T cells by HIV is dependent upon the targeted proteolysis of IRF3 that occurs through a virus-directed mechanism that results in suppression of innate immune defenses. However, the mechanisms by which HIV controls innate immune signaling and IRF3 function are not defined. Here, we examined the innate immune response induced by HIV strains identified through their differential control of PRR signaling. We identified viruses that, unlike typical circulating HIV strains,

lack the ability to degrade IRF3. Our studies show that IRF3 regulation maps specifically to the HIV accessory protein Vpu. We define a molecular interaction between Vpu and IRF3 that redirects IRF3 to the endolysosome for proteolytic degradation, thus allowing HIV Silmitasertib nmr to avoid the innate antiviral immune response. Our studies reveal that Vpu is an important IRF3

regulator that supports acute HIV infection through innate immune suppression. These observations define the Vpu-IRF3 interface as a novel target for therapeutic strategies aimed at enhancing the immune response to HIV.”
“The purpose of this study was to identify brain atrophy specific for dementia with Lewy bodies (DLB) and to evaluate the discriminatory performance of HKI 272 this specific atrophy between DLB and Alzheimer’s disease (AD).

We retrospectively reviewed 60 DLB and 30 AD patients who had undergone 3D T1-weighted MRI. We randomly divided the DLB patients into two equal groups (A and B). First, we obtained a target volume of interest (VOI) for DLB-specific atrophy using correlation analysis of the percentage rate of significant whole white matter (WM) atrophy calculated using the Voxel-based Specific Regional Analysis System for Alzheimer’s Disease (VSRAD) based on statistical parametric mapping 8 (SPM8) plus diffeomorphic anatomic

registration through exponentiated Lie algebra, with segmented WM images in group A. We then evaluated the usefulness of this target VOI for discriminating the remaining 30 DLB patients in group B from the 30 AD patients. Z score values in this target VOI obtained from VSRAD were used as the determinant Carteolol HCl in receiver operating characteristic (ROC) analysis.

Specific target VOIs for DLB were determined in the right-side dominant dorsal midbrain, right-side dominant dorsal pons, and bilateral cerebellum. ROC analysis revealed that the target VOI limited to the midbrain exhibited the highest area under the ROC curves of 0.75.

DLB patients showed specific atrophy in the midbrain, pons, and cerebellum. Midbrain atrophy demonstrated the highest power for discriminating DLB and AD. This approach may be useful for determining the contributions of DLB and AD pathologies to the dementia syndrome.”
“The human respiratory syncytial virus (HRSV) genome is composed of a negative-sense single-stranded RNA that is tightly associated with the nucleoprotein (N).


“Frailty is a common occurrence in


“Frailty is a common occurrence in see more elderly persons and is present

in approximately half of the patients being screened for transcatheter aortic valve replacement (TAVR) therapy. Accurate assessment of the likelihood of benefit from intervention in the older patient with aortic stenosis is critical with both surgical aortic valve replacement and TAVR now available. Whereas risk algorithms are available that are helpful in predicting outcomes in patients undergoing surgical procedures, measures of frailty are not included in the algorithms. When considering intervention in the elderly patient, the addition of frailty assessment to determine the true risk in this population is essential to determine potential benefit. Gait speed as determined by

the 5-m walk test is the most commonly used single test objective measurement of frailty in patients undergoing cardiac surgery and is an independent predictor of mortality and major morbidity. Wider application of this and other objective measures of frailty in the population undergoing TAVR is necessary to determine whether it is predictive in this population also. (J Thorac Cardiovasc Surg 2013;145:S7-10)”
“A 2-layer symbolic network model based on the equilibrium equations of the Rescorla-Wagner model (Danks, 2003) is proposed. The study first presents click here 2 experiments in Serbian, which reveal for sentential reading the inflectional paradigmatic effects previously observed by Milin, Filipovic Durdevic, and Moscoso del Prado Martin (2009) for unprimed lexical decision. The empirical results are successfully modeled without having to assume separate representations for inflections or data structures such as inflectional paradigms. In the next

step, the same naive discriminative learning approach is pitted against a wide range Baricitinib of effects documented in the morphological processing literature. Frequency effects for complex words as well as for phrases (Arnon & Snider, 2010) emerge in the model without the presence of whole-word or whole-phrase representations. Family size effects (Moscoso del Prado Martin, Bertram, Haikio, Schreuder, & Baayen, 2004; Schreuder & Baayen, 1997) emerge in the simulations across simple words, derived words, and compounds, without derived words or compounds being represented as such. It is shown that for pseudo-derived words no special morpho-orthographic segmentation mechanism, as posited by Rastle, Davis, and New (2004), is required. The model also replicates the finding of Plag and Baayen (2009) that, on average, words with more productive affixes elicit longer response latencies; at the same time, it predicts that productive affixes afford faster response latencies for new words. English phrasal paradigmatic effects modulating isolated word reading are reported and modeled, showing that the paradigmatic effects characterizing Serbian case inflection have crosslinguistic scope.

On the stop-signal test, Go reaction time and stop-signal reactio

On the stop-signal test, Go reaction time and stop-signal reaction time were significantly slower in the alcohol-dependent group, compared with healthy controls. Healthy controls slowed their responding after successful and failed stop trials. Slowing after failed stop trials was significantly attenuated in the alcohol-dependent subjects. Go reaction time and post-error slowing were correlated

with chronicity and severity, respectively, in the alcohol-dependent subjects. Problem gamblers did not differ significantly from controls on the stop-signal test, despite trait elevations in impulsivity ratings.

Inhibitory control is impaired in alcohol dependence but occurs in the context of psychomotor slowing.

In addition, alcohol-dependent individuals failed to show behavioral adjustment following failed stops. These deficits Acalabrutinib concentration may represent direct effects of chronic alcohol administration on fronto-striatal circuitry.”
“PKM zeta is an autonomously active, atypical protein kinase C (aPKC) isoform that is both necessary and sufficient for maintaining long-term potentiation (LTP) and long-term memory. The myristoylated zeta-pseudosubstrate peptide. ZIP, potently inhibits PKM zeta biochemically in vitro, within cultured cells, and within neurons in hippocampal slices, and reverses LTP maintenance and erases long-term memory storage. A recent study (Wu-Zhang Selleckchem ATM Kinase Inhibitor et al., 2012), however, suggested ZIP was not effective on a PKM zeta

fusion protein overexpressed in cultured cells. Chelerythrine, a redox-sensitive PKC inhibitor that inhibits PKM zeta and disrupts LTP maintenance and memory storage, was also reported by Wu-Zhang et al. (2012) not to inhibit the expressed PKM zeta fusion protein. However, the efficacy of inhibitors on endogenous enzymes in cells may not be adequately assessed in expression systems in which levels of expression of exogenous enzymes greatly exceed those of endogenous enzymes. Thus, we show, biochemically, that when PKM zeta reaches a level beyond that necessary for substrate phosphorylation such that much of the enzyme is excess or ‘spare’ kinase, ZIP and chelerythrine do not effectively Galactosylceramidase block substrate phosphorylation. We also show that the cellular overexpression techniques used by Wu-Zhang et al. (2012) increase kinase levels similar to 30-40 fold above normal levels in transfected cells. Using a mathematical model we show that at such level of overexpression, standard concentrations of inhibitor should have no noticeable effect. Furthermore, we demonstrate the standard concentrations of ZIP, but not scrambled ZIP, inhibit the ability of PKM zeta to potentiate AMPAR responses at postsynaptic sites, the physiological function of the kinase. Wu-Zhang et al.