As a significant number of stroke victims exhibit cognitive impairment, neuropsychological assessments can enhance poststroke management by identifying pertinent cognitive sequelae and providing salient care recommendations. However, due to operational differences between traditional neuropsychology and stroke services, neuropsychological assessments often remain underutilized in stroke care. We developed a novel care model that incorporated neuropsychological testing into a comprehensive stroke program using the modified vascular cognitive impairment (VCI) half-hour assessment protocol proposed by the National Institute of Neurological Disorders and Stroke—Canadian Stroke Network (NINDS-CSN). The test batteries were administered during the patients' acute admissions and then again upon follow-up in the multidisciplinary stroke clinic. Patient and provider satisfaction was then evaluated. Surveys revealed high provider satisfaction with improved clinic efficacy, improved data turnaround time, and with value neuropsychology services added to the comprehensive stroke program. Results from the 18-item industry standard Press-Ganey surveys showed all scores above 4.4/5.0 for patient satisfaction. This clinic garnered high provider and patient satisfaction after the first year. The (modified) NINDS-CSN VCI assessment protocol demonstrated clinical feasibility, suggestive of an efficient method of providing focused neuropsychological services in a clinical setting that otherwise prohibits traditional, comprehensive cognitive assessments. Stroke is one of the leading causes of long-term disability in the world, and in the United States, an average of one stroke occurs every 40 seconds. By this estimate, 795,000 new or recurrent strokes occur each year, with annual direct and indirect cost of cardiovascular disease and stroke in the United States being estimated at $312.6 billion  found thThe cognitive impairments associated with stroke are heterogeneous , dependiThese points highlight the importance of proactively monitoring stroke patients' cognitive, psychiatric, and functional outcomes during outpatient follow-up. Indeed, a “holistic, comprehensive, interactive approach of an interdisciplinary team” has been regarded as the “hallmark of stroke rehabilitation” . In suchHowever, there is scant literature addressing the feasibility of multidisciplinary stroke care models incorporating cognitive, neurobehavioral, and neuropsychiatric outcomes. In the available literature, it is clear that (a) neuropsychological assessments remain underutilized in pertinent settings and (b) there is little standardization in the methodology of such assessments. In a recent review of stroke research conducted from January 2000 to October 2011, only 488 of the 8826 relevant studies (6%) included cognitive or mood assessment scales . Within Moreover, clinical services that hope to integrate neuropsychological assessment into poststroke care may find its practical implementation challenging. Given the time-sensitive nature of stroke, timely data transfer between providers is crucial not only in achieving the highest patient care standards, but also in promoting patient and provider satisfaction. However, due to operational incompatibilities between traditional neuropsychology clinics and stroke neurology services in most settings, routine neuropsychological assessments are rarely available in the acute poststroke period, and outpatient follow-up appointments are often substantially delayed. These factors often limit provision of neuropsychological assessment in poststroke care, and timely data transfer often remains an ideal rather than a practical, achievable standard.At our tertiary care center, inpatient neuropsychological assessment services were not regularly available for stroke patients, and the outpatient referral system for neuropsychological testing was cumbersome. Patients' quantitative cognitive performance data from neuropsychological testing were seldom available at the time of their stroke clinic posthospitalization follow-up neurology appointments. These systemic barriers brought to our attention the need to implement a streamlined, interdisciplinary model of care that directly incorporated neuropsychology into standard poststroke care.This project aimed to enhance patient services by implementing a multidisciplinary model of poststroke care that more closely integrated neuropsychology and neurology services. We anticipated that such a service would enhance acute poststroke care by (a) making inpatient neuropsychological services part of the standard of care. This would allow for tailored discharge recommendations based on efficient, systematic examination of patients' poststroke cognitive limitations. We also predicated that the benefit of the model would extend to outpatient follow-up practice by (b) providing quantitative measurements of poststroke cognitive recovery, (c) improving patient satisfaction with their poststroke care by mitigating logistical burdens, and (d) improving provider satisfaction with the neurology-neuropsychology collaboration by promoting direct communication and rapid data transfer between providers. We aimed to successfully develop and implement this care model within a 12-month timeframe, thus (e) providing preliminary data regarding the tolerability, feasibility, and utility of the neuropsychological assessment protocol for vascular cognitive impairment proposed by the international harmonization standards for poststroke care endorsed by the National Institute of Neurological Disorders and Stroke and the Canadian Stroke Network  . Financial support for the multidisciplinary stroke service was provided both through the development of a postdoctoral fellowship in stroke neuropsychology, funded by UWHC, and provision of the Ambulatory Care Innovation Grant, an in-house quality improvement program funded by Physicians Plus Insurance Company and the University of Wisconsin Medical Foundation. Total cost for the project, including support of the stroke neuropsychology postdoctoral fellow and purchase of required materials, was under $100,000. After initial implementation, the neuropsychology section of the stroke multidisciplinary team became self-sustaining with service-based revenues.To incorporate neuropsychology into inpatient poststroke care, the stroke neuropsychology postdoctoral fellow was embedded within the inpatient stroke service's morning rounds on two mornings each week. Patients were identified as appropriate for neuropsychological testing on the basis of the fellow's clinical observations, in cooperation with the attending or resident neurologist, occupational therapist, speech-language pathologist, and nurses, who made rounds with the neurology team. Patients so identified were then administered the half-hour NINDS-CSN VCI neuropsychological assessment protocol. Results from this brief inpatient battery; accompanied by salient recommendations for patient supervision, return to school, work, and driving, and medicolegal decision-making capacity, among other considerations; were provided to the stroke team verbally and then in report form via the hospital's electronic medical record system. These recommendations were incorporated into the patient's discharge plan at the discretion of the attending stroke neurologist.As part of the transition of care from discharge to outpatient follow-up, stroke patients were engaged in communication and treatment planning in two ways. First, patients were engaged in a three-way conference call between the inpatient unit clerk and the stroke program coordinator in order to schedule the multidisciplinary poststroke follow-up appointment, and this appointment was included in the inpatient stroke service discharge instructions. Further, informational resources, including letters, brochures, and instructions, were mailed to patients in order to again provide a brief description of the follow-up appointment, the nature of stroke, the purpose and nature of neuropsychological assessment, and resources for additional care.Upon discharge from the inpatient stroke service, an outpatient follow-up appointment was scheduled for each patient for 8 to 12 weeks from discharge during neurology clinic time designated for multidisciplinary stroke services. Each of these follow-up appointments was scheduled as a 2-hour block, with neuropsychological assessment scheduled 1 hour prior to the patient's follow-up appointment with the stroke neurologist. During the neuropsychology portion of the visit, a brief interval history was taken via clinical interview with the patient and any available family members or caregivers, and the NINDS-CSN VCI protocol was repeated in order to provide quantitative measurements of interval cognitive change. Scoring and preliminary interpretation of the test data were performed on-site in the clinic. Results were then provided immediately to the attending neurologist prior to the neurology appointment time, using a side-by-side graphical representation of Time 1 and Time 2 test results. The neurologist then incorporated the results of the neuropsychological testing in their counseling and treatment plan for the patient, often focusing on issues related to capacity for independent living and return to usual activities, such as work and driving.After a pilot year, modified Press-Ganey scales, industry-standard 18-item surveys, were used to assess patient satisfaction with the multidisciplinary stroke clinic model. Specifically, the Press-Ganey scales assessed patient satisfaction with the clinic's appointment logistics, including the scheduling process, access to providers, and communication with care providers. In addition, a 10-item survey assessing stroke neurologists' satisfaction with the new multidisciplinary clinic was administered 12 months after implementation. This survey evaluated providers' satisfaction with clinic scheduling procedures both before and after implementation of the multidisciplinary clinic, data turnaround time both before and after clinic implementation, perceived value of the service to patients, perceived patient satisfaction with their care, and estimation of the clinic's added value to the comprehensive stroke program as a whole. Institutional review board was consulted, and deidentified data use was authorized given that the nature of the data assessed clinical quality improvement.The first goal of the multidisciplinary stroke clinic project was to integrate neuropsychology into the standard of care for stroke patients in our care center. During the 12-month pilot implementation of the multidisciplinary stroke clinic, 114 patients were seen for inpatient cognitive evaluations.Second, we hoped to extend the benefit of neuropsychology's integration to outpatient follow-up care by improving patient satisfaction with poststroke care, in part, by mitigating logistical burdens and improving provider satisfaction with the neurology-neuropsychology collaboration by promoting direct communication and rapid data transfer between providers.Of the 114 stroke patients seen for inpatient cognitive evaluations, 67 patients returned for outpatient follow-up in the multidisciplinary stroke clinic. Remaining 47 patients initially seen were not followed up for multiple clinical reasons, for example, death, out of state residence, being cognitively normal deemed at initial evaluation prior to discharge, or being profoundly impaired with no clinical need for further evaluation. Provision of this service required 67 appointment slots in the multidisciplinary stroke clinic, rather than requiring 134 (67 × 2) separate time slots divided between the outpatient neuropsychology and stroke neurology services. This scheduling benefit required patients to travel to the hospital once for their follow-up appointments rather than making separate trips for neuropsychology and stroke neurology appointments. This represents significant mitigation of the logistical burden on patients, reducing travel costs and increasing feasibility for significantly disabled patients and their caregivers.Prior to the implementation of the multidisciplinary stroke clinic, patients' mean wait time for a poststroke neuropsychological assessment was 5 to 10 months after their discharge from inpatient care. This delay was reduced from 1 to 3 months after implementation of the multidisciplinary stroke clinic, decreasing mean wait time by 7.26 months, a 78.32% decrease in patient waiting time. The 1 to 3 months wait time was also deliberately implemented to gauge poststroke cognitive recovery correlating with the first 90 days of anticipated poststroke improvement.N = 16). This represented a 38% survey response rate.Survey results indicated exceptionally high patient satisfaction with the multidisciplinary stroke clinic, as measured via the Press-Ganey scales. Composite responses to the Press-Ganey scales indicated exceptional patient satisfaction with all domains of service within the multidisciplinary stroke clinic, including logistics and scheduling, patient/provider interactions, patients' perceptions of care quality, and patient confidence in providers. As measured on a self-report Likert scale ranging from 0 to 5, with 0 representing the worst possible rating and 5 representing the best possible rating, patients rated their multidisciplinary stroke clinic care experience in all Press-Ganey assessed domains as at least a 4 out of 5 (range = 4.36 to 5.00). To review patient satisfaction ratings for all domains, see Responses to a 10-item survey of provider satisfaction indicated high provider satisfaction across domains assessed, including providers' perception of improved efficacy of clinic operations and interdepartmental data transfer time after implementation of the multidisciplinary stroke clinic model. Provider survey results also indicated high provider satisfaction with neuropsychology's added value to the comprehensive stroke program. Again, providers rated their satisfaction with the multidisciplinary stroke clinic care model in all assessed domains as at least a 4 out of 5 (range = 4.25 to 4.75). To review provider satisfaction ratings, see Stroke neurologists noted practical benefit from the incorporation of neuropsychology into post-troke services, based on enhanced triage capacities, efficient communication between providers, and rapid data transfer regarding patient outcomes. As neuropsychology was directly embedded into the comprehensive stroke service, and the stroke neuropsychology fellow was available to provide direct input into patient referrals, making it more likely to choose those patients who could best tolerate the NINDS-CSN VCI battery and whose neuropsychological data would likely augment clinical decision making. Effective communication was greatly facilitated, as providers had in-person contact during rounds two mornings a week and in multidisciplinary clinic one afternoon weekly. In addition, for the patients served during the 12-month pilot program, both inpatient and outpatient neuropsychological evaluation data were available during patients' acute stroke admissions and outpatient follow-up appointments. This allowed providers to make recommendations regarding return to school, work, driving, and other usual activities—at least in part—based on quantitative measurements of patients' cognitive capacities rather than relying on clinical judgment alone.The stroke neuropsychology fellow and attending neuropsychologists commented favorably on the general clinical utility of the NINDS-CSN VCI half-hour protocol. First, the protocol could be completed in approximately one hour by most patients, including a focused clinical interview. In addition, the protocol was easily customized, if necessary, in order to individualize assessments as needed without unduly taxing stroke patients' limited tolerance for testing. Examiners did note that patients with significant expressive and receptive aphasias and hemiplegia/hemiparesis involving their dominant hands were often quite limited in their capacity to complete the battery, which heavily relies on verbal and written responses. breadth of useful data across cognitive domains, including cognitive efficiency, speech production, basic visuospatial/constructional skills, verbal learning and memory, and executive functioning. Although the depth of this data did not fully address all clinical issues for every patient, it significantly enhanced clinical understanding, thereby allowing neuropsychology providers to recommend other necessary services, including comprehensive neuropsychological assessments, behind-the-wheel driving evaluations, in-home safety evaluations, skilled nursing placements, and guardianship proceedings.Finally, the protocol provided a sufficientPoststroke cognitive impairment has been demonstrated to be a key predictor of patients' functional outcomes  and riskIn this large tertiary care center, the comprehensive stroke program and neuropsychology service collaboratively implemented a multidisciplinary stroke clinic to address the systemic obstacles that hindered effective, collaborative communication between stroke neurology and neuropsychology providers in their poststroke care efforts. As a result, multiple follow-up visits were condensed into one, reducing travel-related and other associated burdens on patients and reducing stroke patients' wait time between neurology and neuropsychology appointments by over 75%. Instead of requiring weeks or months for data transfer between providers, stroke neurology and neuropsychology providers were able to communicate directly regarding patient's cognitive outcomes in a single clinic appointment. After the first implementation year, the multidisciplinary clinic was associated with high satisfaction ratings from both providers and patients.The half-hour neuropsychological assessment protocol for vascular cognitive impairment, recommended by the 2006 NINDS-CSN international harmonization standards for poststroke care , demonstThe multidisciplinary stroke clinic is a low-cost, easily implemented model of care that quickly becomes financially self-sustaining and is generalizable to both poststroke care teams in other hospitals and other healthcare groups that require multiple disciplines to provide patient care in a streamlined, efficient manner. Such significant increases in provider and patient satisfaction, as well as marked improvements in patient wait time and increases in available patient base, may be anticipated in other services that could utilize neuropsychological services on a regular basis, as well as associated disciplines such as physical therapy, occupational therapy, social work, and health psychology.


Telomerase, regulated primarily by the transcription of its catalytic subunit telomerase reverse transcriptase (TERT), is critical for controlling cell proliferation and tissue homeostasis by maintaining telomere length. Although there is a high conservation between human and mouse TERT genes, the regulation of their transcription is significantly different in these two species. Whereas mTERT expression is widely detected in adult mice, hTERT is expressed at extremely low levels in most adult human tissues and cells. As a result, mice do not exhibit telomere-mediated replicative aging, but telomere shortening is a critical factor of human aging and its stabilization is essential for cancer development in humans. The chromatin environment and epigenetic modifications of the hTERT locus, the binding of transcriptional factors to its promoter, and recruitment of nucleosome modifying complexes all play essential roles in restricting its transcription in different cell types. In this review, we will discuss recent progress in understanding the molecular mechanisms of TERT regulation in human and mouse tissues and cells, and during cancer development. Telomeres are the protective caps on chromosomal ends that are required for chromosomal stability and cell survival. Telomeres are replenished by telomerase, a ribonucleoprotein complex containing a catalytic subunit (TERT), an RNA template (TERC), and other associated proteins . TelomerTelomere length regulation and mechanisms of proliferative senescence are not evolutionarily conserved, even among mammals. In a comparative analysis of telomere length and telomerase expression in cells of over 60 mammalian species, Gomes and colleagues concluded that the ancestral mammalian had human-like short telomeres and repressed telomerase expression . Cells iLaboratory mice are the most commonly used animal models for human development, aging, and diseases. While telomere length serves as a critical counting mechanism for cellular senescence in human cells, mice do not exhibit telomere-mediated replicative aging. Compared to humans, telomere homeostasis in mice is distinctive in two ways: Laboratory mice express ubiquitous telomerase activities in somatic tissues and possess long heterogeneous telomeres ,7. ThereMouse models of human diseases have become a central part of biomedical research. Laboratory mice provide the most experimentally accessible mammalian models that share genes, organs, and systemic physiology with humans. However, many mouse models do not comprehensively mimic human disease progression, posing challenges in their exploitation to study human diseases. This may have contributed to the high failure rates of human clinical trials, particularly in oncology, predicating the need for improved preclinical data from mouse models . A princhTERT is a limiting subunit of telomerase in most human tissues. Whereas TERC RNA and other telomerase-associated proteins are expressed in most cell types, hTERT expression is highly regulated and its expression correlated with telomerase activity in many cell types. Studies have indicated that hTERT transcription is the primary step of telomerase regulation . hTERT thTERT expression is stringently regulated in most adult somatic tissues. It was previously shown that the hTERT gene was embedded in a condensed chromatin domain in many somatic cells . This reOne of the bottlenecks in the study of hTERT gene regulation is that the transiently transfected hTERT promoter reporters often do not recapitulate the regulation of endogenous promoter in host cells. For example, transient reporters of the hTERT promoter were similarly active in both telomerase-positive and negative cells ,20. ThisThe lack of somatic hTERT expression, compared to those of mTERT, are most likely resulted from its much stronger repression—in human cells, owing to the genomic sequences of the hTERT locus. Indeed, while ectopic hTERT expression is sufficient for the immortalization of many human cell types , spontanThe order of TERT and its neighboring loci, upstream gene CRR9 and downstream Xtrp gene, is the same in human and mice A. The inTaken together, the interspecies difference of TERT regulation resulting from divergent chromatin environment of hTERT and mTERT loci likely contributes to different telomere length regulation in human and mice. Although primary functions of the TERT protein are evolutionarily conserved, the differential telomerase regulation and telomere length between human and mice have profound impact on the uses of laboratory mice as models of human aging and diseases.While chromatin environment and epigenetic regulation control hTERT regulation during development and cell differentiation, hTERT transcription is also regulated by binding of transcription factors (TFs) to its promoter. The hTERT promoter contains abundant potential TF binding sites and these sites may play important roles in hTERT transcription in response to changes of physiological conditions and during tumorigenesis. Several sequence-specific transcriptional factors have been identified to bind directly to the hTERT promoter, like c-Myc, USFs, Sp1, Ets, E2F1, AP-1 and ER B 2,25]..25]. MosThere are two canonical E-box consensus sites at the hTERT core promoter, located at upstream 165 bp and downstream 45 bp relative to the hTERT transcriptional start site (TSS). These sites are known binding sites for c-Myc super family TF complexes. c-Myc is a proto-oncogene and its overexpression increases telomerase activity by elevating hTERT transcription level in normal human epithelial cells and primary fibroblasts . The repMax dimerization protein 1 (Mad1) was identified as a transcriptional repressor during a screening for hTERT regulators . The expE2F1 promotes cell proliferation and mediates cell apoptosis. There are two non-canonical E2F1 binding sites (E2F) at the hTERT promoter, −174 and −98 (CGCGC) relative to the start of transcription . E2F1 ovTransforming growth factor-β (TGF-β) represses telomerase activity by down-regulating hTERT mRNA level. Snail, one of the downstream targets of TGF-β induction, is highly expressed during embryogenesis and plays a critical role in the stem cell division. Expression of snail was increased in immortalized human cells, in which hTERT transcription was activated . Up-reguHistone acetylation and deacetylation are critical for gene regulation. Trichostatin A (TSA), an inhibitor of classes I and II histone deacetylases (HDACs), increased hTERT mRNA level in normal cells and telomerase-negative immortal cell lines ,42,43. MGenomic sequence around the hTERT promoter is highly GC-rich. Methylation of two CpG islands near the promoter, located at −900 bp and −4000 bp upstream of ATG codon, respectively, was reported to be associated with lower hTERT expression in B-cell lymphocytic leukemia . AberranhTERT expression is induced in cancer cells but absent in most somatic cells. First, transcription activators lead to up-regulation of TERT expression in cancer cells. Increased c-Myc expression in cancer cells resulted in the binding of c-Myc/Max complex to E-boxes at hTERT promoter . Sp1 bonSecond, germline and somatic mutations of the hTERT promoter affect hTERT transcription expression, and impact cancer development. Multiple mutations at the hTERT promoter have been discovered in recent years with the use of next-generation sequencing techniques . Two hotGenome-wide association studies revealed that SNP sites at the chromosomal region 5p15.33, where hTERT gene located, were associated with increasing cancer risks . SNPs wiIn addition to point mutations, chromosomal rearrangement and amplification involving the hTERT locus have been discovered in immortal cells and cancer cells. In our studies, the chromosomal breakage points have been mapped to the upstream regions of hTERT promoter in telomerase positive immortal cells but not in the telomerase negative cells and their parent cells . The levhTERT expression strictly limits telomerase activation in most of somatic cells, whereas mTERT expression is detectable in most of mouse tissue cells. The interspecies differences between human and mice suggest an improved mouse line, in which both telomerase regulation and telomere length controls are humanized, would considerably benefit the studies of human aging and cancer using mouse models.


In many systems, sleep plays a vital role in memory consolidation and synaptic homeostasis. These processes together help store information of biological significance and reset synaptic circuits to facilitate acquisition of information in the future. In this review, we describe recent evidence of sleep-dependent changes in olfactory system structure and function which contribute to odor memory and perception. During slow-wave sleep, the piriform cortex becomes hypo-responsive to odor stimulation and instead displays sharp-wave activity similar to that observed within the hippocampal formation. Furthermore, the functional connectivity between the piriform cortex and other cortical and limbic regions is enhanced during slow-wave sleep compared to waking. This combination of conditions may allow odor memory consolidation to occur during a state of reduced external interference and facilitate association of odor memories with stored hedonic and contextual cues. Evidence consistent with sleep-dependent odor replay within olfactory cortical circuits is presented. These data suggest that both the strength and precision of odor memories is sleep-dependent. The work further emphasizes the critical role of synaptic plasticity and memory in not only odor memory but also basic odor perception. The work also suggests a possible link between sleep disturbances that are frequently co-morbid with a wide range of pathologies including Alzheimer’s disease, schizophrenia and depression and the known olfactory impairments associated with those disorders. The olfactory system Figure  is remarTogether, these experience-induced changes can modify olfactory system sensitivity and acuity, as well as link odor quality to associative meaning or hedonic valence. The changes can last as short as a few seconds, in the case of short-term odor adaptation  can occur for motor skill memories, declarative/episodic memories, emotional memories and sensory memories/perceptual learning  or paradoxical sleep and slow wave sleep (SWS), also known as non-paradoxical or NREM sleep Figure . In the The predominant oscillatory activity in SWS is in the δ-band (0.5–4.0 Hz), which includes the less than 1 Hz slow oscillation Buzsáki, . These oA special type of oscillation seen in the hippocampus , which gradually decreases from early to late sleep Borbély, . HoweverAs noted above, sleep-dependent consolidation has been demonstrated for a variety of forms of information including declarative/episodic, motor, emotional and sensory memory. The prevailing view is that during sleep, recently acquired information is replayed, both within local neural networks  and across networks . This replay allows recently strengthened synapses to solidify those changes either biochemically or structurally, while other less critical synapses are reset to basal strengths, ready for plasticity another day  does not transfer to judgments of diagonal or horizontal lines. Similar selective improvements can be made in the other thalamocortical senses with appropriate training. The learned enhancement in sensory acuity is associated with narrowed receptive fields in sensory neocortex and/or enlargement of the sensory cortical region devoted to that segment of the sensory world . ACh plays a major role in odor processing and plasticity throughout the olfactory pathway from the OB  odor experience that occurred during fast-wave state Wilson, . PiriforOdor fear conditioning can induce either highly odor-specific fear responses  or more generalized odor-evoked freezing, depending on the nature of the training protocol (Chen et al., To test this, we utilized olfactomimetic electrical stimulation of the OB (Mouly et al., Rats were trained using these olfactomimetic stimuli in a differential fear conditioning task, and then placed in a quiet chamber for the 4 h post-training. Piriform cortical local field potentials and neck muscle EMG were recorded to identify sleep/wake states. Animals that were trained and then left alone during the 4 h post-training period displayed CS+ specific freezing the following day. However, the strength and accuracy of this memory could be modulated by post-training CS+ imposed replay (Barnes and Wilson, We hypothesize that during post-training slow-wave sleep, ensembles of piriform cortical neurons that had been recently co-activated during training, preferentially fire together during piriform cortical sharp-waves in a form of spontaneous replay. This co-activation is facilitated by a release of intracortical association fiber synapse suppression due to low levels of cholinergic activity during slow-wave sleep (Hasselmo and Bower, B receptor agonist baclofen bilaterally into the piriform cortex during the post-training period. Baclofen selectively suppresses association fiber synapses in the piriform cortex (Tang and Hasselmo, If this model is correct, then disrupting intracortical association fiber synapses during the post-training period should impair odor memory strength and/or memory precision. To test this, we infused the GABATogether, this work suggests that piriform cortical sharp-wave activity during post-training slow-wave sleep allows for a strengthening of ensemble representation of the learned odor. This post-training replay can modify both the strength and the precision of the odor memory, at least in part via plasticity within the piriform cortex itself. However, it must be noted that co-activation of large piriform cortical ensembles during sharp-waves also result in propagation of spike trains out of the cortex to its monosynaptic neighbors. Thus, this provides an opportunity for piriform cortical manipulation not only of intra-cortical synapses, but also of its efferent targets during slow-wave sleep. These “downstream” effects have been the focus of recent work in Kensaku Mori’s group (Manabe et al., One important role for odor learning is for memory of novel flavors. The majority of the perception of flavor is derived from volatile food odorants delivered to the olfactory epithelium via retronasal smell. While humans are believed to have the most advanced retronasal olfactory abilities (Shepherd, As noted above, odor memory is associated with changes throughout the olfactory pathway. Perhaps the most extreme neural correlate of odor memory is differential survival of OB granule cells. OB granule cells are inhibitory interneurons which modulate the excitability of OB mitral and tufted cells and are the primary target of descending inputs from olfactory cortex. Importantly, granule cells display adult neurogenesis Bayer, , with suMori’s group suggests that the strong descending sharpwave-associated pyramidal cell spiking from the olfactory cortex to the OB during slow-wave sleep (Manabe et al., Combining the findings from these two paradigms leads to the suggestion that post-odor exposure slow-wave sleep contributes to changes across the olfactory system that contribute to both the strength and precision of odor memory Figure . Slow-waOlfactory deficits are associated with a variety of disorders including, but not limited to Alzheimer’s Disease Murphy, , ParkinsThus, we speculate that a contributing factor to the widespread occurrence of olfactory disorders, particularly odor identification, across diverse pathologies may be related to underlying sleep disorders. Memory and neural plasticity are integral not only to odor memory, but also to basic odor perception (Wilson and Stevenson,


The stiffness of the myogenic stem cell microenvironment markedly influences the ability to regenerate tissue. We studied the effect of damaged myofibers on myogenic progenitor cell (MPC) proliferation and determined whether the structural integrity of the microenvironment contributes to phenotypic changes.Individual myofibers were isolated and cultured for 6 days. During this period, the cytoskeleton of myofibers and transcription factors regulating MPC differentiation were characterized by immunostaining. Atomic Force Microscopy (AFM) was performed to measure stiffness of cultured myofibers. Healthy and damaged myofibers, and their associated MPCs, were studied in skeletal muscle from dystrophic and tenotomy mouse models. MPCs were cultured on stiffness-tunable substrates, and their phenotypes were assessed by immunostaining of myogenic transcription factors.ex vivo starting from day 1 and that this is associated with a marked increase in the number of proliferative MPCs (Pax7+MyoD+). The myofibers collapsed due to a loss of viability as shown by Evans blue dye uptake and the disorganization of their cytoskeletons. Interestingly, collapsed myofibers in mdx skeletal muscles were similar to damaged myofibers in that they lose their viability, have a disorganized cytoskeleton (actin and α-actinin), and display local MPC (MyoD+) proliferation at their periphery. In a tenotomy model that causes loss of muscle tension, the cytoskeletal disorganization of myofibers also correlated with the activation/proliferation of MPCs. A deeper analysis of collapsed myofibers revealed that they produce trophic factors that influence MPC proliferation. In addition, collapsed myofibers expressed several genes related to the basal lamina. Immunostaining revealed the presence of fibronectin in the basal lamina and the cytoplasm of damaged myofibers. Lastly, using atomic force microscopy (AFM), we showed that collapsed myofibers exhibit greater stiffness than intact myofibers. Growing MPCs on a 2-kPa polyacrylamide-based substrate, exempt of additional microenvironmental cues, recapitulated proliferation and reduced spontaneous differentiation compared to growth on a 0.5-kPa substrate.We showed that individual myofibers tend to shrink or collapse when cultured Our results support the notion that collapsed or damaged myofibers increase the structural stiffness of the satellite cell microenvironment, which in addition to other cues such as trophic factors and changes in extracellular matrix composition, promotes the proliferation and maintenance of MPCs, required for myofiber repair.The online version of this article (doi:10.1186/s13395-015-0030-1) contains supplementary material, which is available to authorized users. Satellite cells are quiescent muscle stem cells that play a determinant role in myofiber turnover and postnatal regeneration . Satelliin vitro myogenic stem cell renewal, proliferation, differentiation, and regenerative potential thymidine incorporation showing that TA tenotomy promotes the proliferation of stromal and myogenic cells [mdx mice in which MPCs proliferate locally around EBD+ myofibers, there were virtually no EBD+ myofibers in the tenotomized muscles, and the MPCs were rather scattered throughout the tissue, with a few clusters of MPCs. Despite the absence of degenerating EBD+ myofibers, the vast majority of myofibers displayed generalized disorganization, as shown by the loss of width uniformity. In addition, F-actin and α-actinin staining revealed that most of the myofibers had a disorganized cytoskeleton as shown by the lack of periodicity. As such, in addition to collapsed necrotic myofibers, the physiological loss of myofiber tension in vivo appeared to be sufficient to induce the activation and proliferation of MPCs in vivo.Since myofiber collapse or shortening appeared to be a key for MPC activation/proliferation, we determined whether MPC proliferation is affected by traumatic muscle shortening. We used a tenotomy model to show that the number of MPCs is significantly higher in tenotomized than in contralateral control muscles. This increase in MPC proliferation confirmed previous findings with respect to [ic cells . Unlike in vitro. In fact, IGF-1 expression was higher in collapsed myofibers, which corroborated previous reports that IGF-1 secretion increases in differentiated skeletal muscle cells undergoing biomechanical stimulation (stretching) [in vitro and in vivo. Altogether, our findings highlight the complex nature of the interactions between biochemical factors and the ECM in the MPC microenvironment. This was especially true for MPCs cultured on plastic petri dishes, which are dissociated from their biochemical and biomechanical microenvironments and which invariably lose their stemness and regenerative potential [Changes in the satellite cell niche microenvironment directly impact MPC activities ,3,43. Thetching) . Furtheretching) . Interesetching)  and muscetching) ,66, was otential ,20.A previous study using AFM-based indentation experiments to determine the stiffness value of skeletal muscle reported a Young’s modulus of approximately 12 kPa . However6-kPa substrates, which correspond to brain, cartilage, and plastic petri dish stiffnesses, respectively [Interestingly, our AFM-based experiments with individual myofibers revealed that the stiffness values of intact myofibers are four times lower than those of collapsed myofibers (0.5 versus 2.0 kPa). When the stiffness values were recapitulated in gel substrates, the 2-kPa environment promoted the proliferation of MPCs and inhibited their differentiation, maintaining MPCs in a proliferative state. This was in agreement with previous work showing that MPC proliferation is strongly influenced by elasticity while their differentiation is modulated by the protein coating . Studyinectively . While sOur results provided new evidence that minor biomechanical alterations in the microenvironment, such as stiffness, are sufficient to directly impact the proliferative and differentiation capacities of muscle stem cells. In essence, they reinforce the role of microenvironmental cues such as mechanical constraints and local factors in MPC behavior during physiological muscle repair, muscle regenerative disorders, and aging.


Eleusine coracana (L.) Gaertn.] is a crop with potentially tremendous but under-explored source of nutraceutical properties as compared to other regularly consumed cereals. In the era of growing divide and drawback of nutritional security, these characteristics must be harnessed to develop finger millet as a novel functional food. In addition, introgression of these traits into other staple crops can improve the well-being of the general population on a global scale. The objective of this review is to emphasize the importance of biofortification of finger millet in context of universal health and nutritional crisis. We have specifically highlighted the role that recent biotechnological advancements have to offer for enrichment of its nutritional value and how these developments can commission to the field of nutritional biology by opening new avenues for future research.The science of nutritional biology has progressed extensively over the last decade to develop food-based nutraceuticals as a form of highly personalized medicine or therapeutic agent. Finger millet [ Eleusine coracana (L.) Gaertn.], possessing wholesome traits to meet global nutritional infirmity, but also explores its nascent significance as means of advancing the future of nutraceutical industry.The World Health Organization (WHO) recognizes that maintenance of optimum global public health requires not only prevention of early onset of chronic diseases but also promotion of a healthier aging and ultimately prolongation of life. Despite its tremendous advancement, medical science still faces a dual challenge to promote public health: side effects of medicines and their economic encumbrance. Going back to a well-recognized dogma of ancient Indian Siddha literature and Hippocrates ‘food is medicine and medicine is food,’ diet and dietary habits have been established to play important role for exalted health benefits based on different properties of foods . Thus, gThe health and nutrition-related disorders in various populations have been correlated to the differential interaction of nutrient-dependent genetic diversity . ResearcThe scientific area of ‘nutrigenomics’ is actually integration of genomics into nutrition sciences. It uses functional genomic tools for genome-wide identification of genes which are influenced by food components, resulting in alterations in homeostatic pathways in a biological system. It is a challenging area as a single nutrient can have different targets, affinities or specificities based on population under study. Quantitative techniques for integrated or independent study of nutrients on subtle changes in gene expression (transcriptomics), proteins (proteomics), or metabolites (metabolomics) in single cell as well as whole organism have increased the knowledge about nutrient-gene interaction aiding nutritional research tremendously. With the completion of several genome-sequencing projects, new genes involved in several metabolic pathways have been identified. Microarray analyses in nutrigenomic studies have defined gene expression pattern changes for 1000s of such genes . DietaryIndividual preferences and differences in diet and lifestyle also affect the genetic makeup and such food-responsive gene-based variation can be understood by nutrigenetics. Conversely, it also aims to study that how genic variations such as single nucleotide polymorphisms coordinate individualistic responses to diet in the whole population. It is driven by the hypothesis of selection of genes based on their ‘suspected’ effect on health, for example, insulin signaling genes for diabetes or lipoprotein genes as candidates for obesity. Nutrigenetics can reveal polymorphism in response to food components in genetically diverse population and identify groups which could be predisposed to nutrition-relation disorders or those which could gain substantial benefit from dietary intervention. Thus, nutrigenomics and nutrigenetics research needs to be coupled with a number of other academic fields like molecular biology, molecular nutrition, pharmacogenomics, and molecular medicine for a customized designing of knowledge-based functional foods on a large scale.Pennisetum glaucum (L.) R. Br.], finger millet [E. coracana (L.) Gaertn.], foxtail millet [Setaria italica (L.) P. Beauvois.], and proso millet [Panicum miliaceum (L.)]. They are considered as a storehouse of nutritional properties and remedy for malnutrition that affects a vast majority of populations in developing countries . From the nutritional perspective, finger millet is considerably rich in minerals and its micronutrient density is higher than that of the world’s major cereal grains; rice and wheat . However, these ingredients need to be isolated and developed as nutraceuticals and functional food, without disturbing the inherent functional and nutritional traits and in a cost-efficient manner. Nutraceutical properties of finger millet are discussed in details in the following sections.Recent research has shown that diets rich in plant-based foods, particularly whole-grain cereals, are protective against several degenerative diseases such as diabetes, cardiovascular diseases, few types of cancers, metabolic syndrome and Parkinson’s disease . In spitp-hydroxybenzoic acid, vanillic acid, syringic acid) while rest of the fraction were either cinnamic acid derivatives  or flavonoids  could be indirectly connected with its ability to alleviate type-2 diabetes risk. It has been reported that consumption of multigrain flour containing 30% finger millet proportion significantly lowered the plasma glucose levels  or because of mutual endurance to insulin secretion and action (type-2). Clinically, -prandial hyperglycemia can be managed by chemical synthetic inhibitors of α-glucosidase and pancreatic amylase, the potential safety of natural inhibitors is overt. Phenolic extracts from finger millet seed coats were shown to be effective inhibitors of these enzymes . This pre levels , which ce levels . The ante levels . Additioe levels . Very smications . Therefoin vitro copper-mediated human LDL cholesterol oxidation system, however, this function was varietal or dose-dependent with respect to specific phenolics . Finger millet is low in fat content and is good for obese people. Supplementation with finger millet bran helped to prevent high-fat diet-induced obesity and increased the abundance of beneficial gut microflora in rodent models  and obesity (excessive dietary consumption). Being a highly balanced and easily digestible grain, finger millet presents an ideal diet for both the groups. For the people suffering from protein-energy malnutrition, it is an excellent source carbohydrate (80%) and protein (7–9%) with essential amino acids like valine, methionine, and tryptophan which otherwise are scarcely present in a vegetarian diet. Further, minerals like calcium, phosphorus, potassium, and iron and vitamins like and thiamine, niacin, and riboflavin are also abundant  is predicted to be the uplands of East Africa, especially Ethiopia and Uganda around 5000 years ago from wild finger millet race , randomly amplified polymorphic DNA (RAPD), and inter simple sequence repeat amplification (ISSR) markers were utilized to assess genome origins and genetic diversity among five species of the genus Eleusine . RAPD maEleusine . DiverseEleusine . Genic aEleusine . In a crEleusine . AdditioEleusine . This apEleusine . Babu etEleusine . Thus, gEleusine .The identified molecular markers have promoted the potential to boost future breeding for nutritional improvement of finger millet especially in terms of mapping and tagging important QTLs and construction of genetic maps. The first genetic linkage map of finger millet, constructed at John Innes Centre, UK, covered 1,100 cM of genome across 27 linkage groups (LGs) using 212 RFLP markers . Since, Table 3). High leaf blast disease resistant finger millet transgenics have been successfully produced using biolistic or gene gun method  were the only approaches available for gene expression studies. The situation has changed dramatically during the last few years with the advent of next-generation sequencing and analytical tools . RecentlProteomics is the large-scale study of the complete set of proteins and analysis of their expression, structure and function . Proteomp-hydroxybenzoic, vanillic, and ferulic acids), cinnamic acid ; and quercetin (flavonoid; Metabolomics and ionomics are one of the recent “omics” technologies that allow the analysis of a low molecular weight biomolecules and elemental composition synthesized or consumed by a biological system, respectively. These technologies can provide a direct functional statement of a plant’s nutritive value by determination of its metabolic biochemistry, ionic contents, and phytochemicals . VariatiArabidopsis was developed to identify and capture the key quantitative attributes of plant development and architecture (A multidisciplinary approach which aims to integrate biological sciences with computer studies and mathematics, called as systems biology, endeavors to quantitatively characterize the wealth of transcriptome, proteome, and metabalome datasets and simulate them to forecast an informational pathway . In one itecture . The sysitecture . These wThe growing demand for healthier food, effectiveness and quality of consumed product and increased public and healthcare industry awareness are the major factors that have contributed toward the formation of a nutraceutical market that is envisioned to grow many folds in coming years. The Food and Drug Administration has also released regulations that support this emerging industry therefore fostering scientific research. Hence, an immediate goal should be identification of health benefitting factors to enhance the essential nutrient levels in staple crops to significantly impact human nutrition worldwide. Targeting of nutritionally important genes and proteins through the emerging biotechnology tools and techniques can lead to creation of ‘smart’ biofortified crops. Products from these value-added crops can help to cope with several problems such as protein-energy malnutrition. Research should establish impact of these products on the body’s absorption, defense, regulation of homeostasis and nervous systems, and then delve into hypo-allergenic foods and modern approaches to nutraceutical production. The initial research has shown finger millet to have a bright future in the nutraceutical industry and provides a scientific rationale for its use as an economically viable nutrient store to depreciate chronic pathologies. From the consumer’s perspective, establishment of finger millet as a nutraceutical can surpass the usual wait, efforts and cost inputs to bring conventional healthcare to the market and provide “self-care” for their satisfaction. Additionally, for a global scale, exploitation of its rich nutritional value assumes importance to provide food security, agricultural development, self-dependence and economic enhancement of developing countries. With the increasing knowledge about the nutraceutical properties of finger millet, the day is not far when this crop and its various products will find their place in the day to day menu of every individual.AK conceptualized the manuscript. MM, SK, AKG, Sadhna Singh, MS wrote the manuscript. SG, BKB, Salej Sood assisted and edited the manuscript. SP and RSY contributed in critically revising the draft and updating the manuscript for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Stem Cell Research & Therapy, Qin et al. demonstrated enhanced therapeutic properties of adipocyte-derived stem cells by genetically engineering them to express periostin.Periostin is a matricellular protein that is reactivated during tissue damage and repair and has been shown to be a critical regulator of multiple biological pathways involved in the repair of tissue after myocardial infarction, peripheral vascular disease, and skin wounds. The tissue repair properties attributed to periostin make it an ideal candidate to enhance the therapeutic properties of donor cells such as mesenchymal stem cells from adipocyte tissue. In a recent article in  Stem Cell Research & Therapy, Qin et al. modified adipose-derived stem cells (ADSCs) to improve their survival [Mesenchymal stromal or stem cells (MSCs) from different tissue sources have a proven track record of enhancing tissue repair through the direct contribution to new tissue by engrafting and differentiating into muscle, endothelial cells, adipocytes, and bone. MSCs also have a powerful paracrine signaling system resulting in the enhancement of endogenous repair mechanisms. Despite the advantages of cell-based therapy, one of the common problems facing the use of cells for tissue repair is the low survival rate of the donor cells. In a recent article in survival . Normallsurvival . There hsurvival –5, but mQin et al. modified ADSCs to express the periostin gene by using lentivirus vectors and determined that the modified ADSCs had superior tissue repair properties compared with unmodified control ADSCs by using a model of peripheral vascular disease (PVD) . PeriostTissue repair, whether in the context of PVD , skin wowww.ClinicalTrials.gov). Previous studies have used non-integrating plasmids to deliver genes [Adipose tissue, bone marrow, and umbilical cord tissues are all commonly used sources of MSCs to treat tissue damage. Qin et al. used ADSCs instead of bone marrow-derived MSCs, cells which share similar properties, because adipose tissue collection is minimally invasive, making them more clinically applicable. Qin et al. also chose lentivirus to introduce the periostin gene into the cells. Although the use of lentivirus faces more stringent safety measures compared with using other non-permanent methods to modify cells, lentivirus vectors offer advantages such as stable transfection of the cells, resulting in long-term periostin production at the site of cell transplantation. Furthermore, lentiviruses do not induce an immune reaction in the host, and lentivirus-modified cells are currently being used in multiple clinical trials  and tumor necrosis factor alpha (TNFα), which in turn regulate critical ECM protein and matricellular proteins, including periostin, heat shock protein (hsp), and fibronectin . Periost


There is increasing global demand for food, bioenergy feedstocks and a wide variety of bio-based products. In response, agriculture has advanced production, but is increasingly depleting soil regulating and supporting ecosystem services. New production systems have emerged, such as no-tillage, that can enhance soil services but may limit yields. Moving forward, agricultural systems must reduce trade-offs between production and soil services. Soil functional zone management (SFZM) is a novel strategy for developing sustainable production systems that attempts to integrate the benefits of conventional, intensive agriculture, and no-tillage. SFZM creates distinct functional zones within crop row and inter-row spaces. By incorporating decimeter-scale spatial and temporal heterogeneity, SFZM attempts to foster greater soil biodiversity and integrate complementary soil processes at the sub-field level. Such integration maximizes soil services by creating zones of ‘active turnover’, optimized for crop growth and yield (provisioning services); and adjacent zones of ‘soil building’, that promote soil structure development, carbon storage, and moisture regulation (regulating and supporting services). These zones allow SFZM to secure existing agricultural productivity while avoiding or minimizing trade-offs with soil ecosystem services. Moreover, the specific properties of SFZM may enable sustainable increases in provisioning services via temporal intensification (expanding the portion of the year during which harvestable crops are grown). We present a conceptual model of ‘virtuous cycles’, illustrating how increases in crop yields within SFZM systems could create self-reinforcing feedback processes with desirable effects, including mitigation of trade-offs between yield maximization and soil ecosystem services. Through the creation of functionally distinct but interacting zones, SFZM may provide a vehicle for optimizing the delivery of multiple goods and services in agricultural systems, allowing sustainable temporal intensification while protecting and enhancing soil functioning. Intensification of agriculture has been vital for increasing global food supply and alleviating hunger for millions of people . In addiOne option for securing the productivity of existing agricultural land while also enhancing delivery of soil ecosystem services is by integrating the high productivity of intensive field crop production systems (including intensive tillage) with the improvements in soil quality associated with stringent limitations on tillage. Herein, we present evidence that a novel approach to management of field crop agroecosystems – soil functional zone management (SFZM) – can promote such integration. As detailed below, SFZM entails the creation and management of distinct yet complementary soil functional zones that have potential to reduce trade-offs between short-term productivity and soil quality.We believe SFZM to be a previously unrecognized strategy for expanding the range of ecosystem service production from field crop agroecosystems. Several forms of SFZM  have been studied extensively in terms of their effects on a range of crop and soil attributes. Here, we expand upon this level of analysis and understanding through a broad exploration of ecosystem service production and underlying agroecological processes in SFZM, drawing on a wide range of evidence and identifying critical knowledge gaps in understanding of SFZM. In our analysis, we focus first on supporting and regulating services, and then examine the potential of SFZM to increase productivity of agricultural systems . In particular, we consider the role of SFZM in supporting temporal intensification, which aims to enhance provisioning services by expanding the annual time period in which harvestable crops are grown. We consider the potential dynamics of agroecosystems under SFZM, and the role of these dynamics in improving the sustainability of temporal intensification. We focus on the dynamic implications of ‘virtuous cycles’ (self-reinforcing feedback processes with desirable effects) that may occur in SFZM. Such feedback processes may serve to reduce trade-offs between provisioning, supporting, and regulating services in temporal intensification.Figure 1). While SFZM does not necessarily involve novel management practices , it provides a novel framework for enhancing ecosystem service production in field crop agroecosystems.Soil functional zone management is a novel concept of field crop agroecosystem management that seeks to create distinct, yet functionally complementary soil zones through non-uniform management of tillage and crop residues. These zones can be tailored for a variety of different functions or ecosystem services and can be permanent or change locations between seasons. At its most basic, SFZM involves a zone of ‘active turnover’, managed to optimize conditions for seed germination and crop growth; and an adjacent ‘soil building’ zone, which is managed to protect soil organic matter (SOM), enhance soil water holding capacity and provide habitats for soil organisms. At present, the two most widely practiced implementations of SFZM are ridge tillage and strip tillage . Despite advances in precision agricultural application of fertilizer and agrochemicals, tillage is still predominantly applied homogeneously , SFZM creates spatial heterogeneity over small (<1 m) spatial scales. Relative to non-zonal tillage, such enhancement of within-field heterogeneity across space and time serves to enhance the range of soil physical conditions and functional biodiversity within a row-crop agroecosystem. Increasing heterogeneity can enhance biodiversity by providing habitat and other key resources to a wider range of organisms. This expansion of resource diversity in space and time can support effective resource partitioning and increased diversity of microhabitats, allowing coexistence of soil organisms and increased functional biodiversity . We hypothesize that these refugia can support faunal and fungal diversity in a similar way to no-tillage, and provide a base from which slow-growing organisms with longer generation times (K-selected organisms) might be able to recolonize disturbed areas. In essence, we propose that SFZM, by expanding both habitat and resources relative to conventional soil management, can enhance both provisioning and soil regulating and supporting services by enhancing soil biodiversity.Conventional soil management is typically characterized by frequent and intense disturbance  combined with low plant resource diversity . These factors lead to reduced abundances and diversity of soil organisms in conventional systems compared with no-tillage and other systems with reduced tillage and more diverse crop rotations . MoreoveSecuring high levels of agricultural production while simultaneously improving regulating and supporting soil ecosystem services requires management strategies that expand the range of service production . As outlThe creation of undisturbed refugia for soil microbiota, particularly filamentous fungi, through targeted disturbance is one pathway by which SFZM may increase the supply of supporting soil services. Such refugia should impact carbon (C), nitrogen (N), and phosphorus (P) cycling to the benefit of above-ground productivity. Nutrient cycling among organic and inorganic pools is driven by microbial turnover, with fungi generally thought to be more effective at storing C and N in organic matter than bacteria , while tr-selected, producing more reproductive spores and fewer soil-exploring hyphae than K-selected AMF . The disturbed region exposes labile organic matter and aerates the soil, providing excellent conditions for nutrient turnover immediately after disturbance  by AMF in SFZM systems  likely enables them to persist in ridge tillage systems .Such refugia may explain enhanced P delivery to maize  Osbeck; Brassica oleracea L.) system (The key to N synchrony may be to manage N supply in both space and time . This is) system . This maDiabrotica virgifera Le Conte) eggs in furrow positions and delayed larval development following an intra-seasonal ridging event. They suggested the ridging event relocated the previously uniformly dispersed eggs into the furrow while also burying the eggs under surface debris, lowering soil temperatures, and slowing larval development ; fungal hyphae play an important role in the formation and stability of soil aggregates . Recent Soil functional zone management may also provide regulating services that contribute to the suppression of weeds. Non-herbicidal weed suppression services will become increasingly valuable as populations of weeds that are resistant to glyphosate and other herbicides continue to become more abundant. The problem of herbicide resistant weeds is especially acute in conventional no-tillage systems, and particularly in those systems that rely on herbicide resistant crops, because of their exclusive reliance on herbicides for weed control . SFZM, tFigure 1). This uncoupling of soil temperatures and residues from areas of soil disturbance allows soil in row positions to warm more rapidly early in spring, while preserving an extensive AMF mycelial network for rapid root colonization in inter-rows (Solanum lycopersicum L.) performance was improved by AMF when in competition with bahiagrass . Therefore, by actively managing the placement of phytotoxic cover crop residues, SFZM can minimize some of the potential trade-offs associated with the use of cover crops. The process of concentrating crop residues also promotes survival of soil pathogens in inter-row positions, by increasing inter-row soil moisture content , can also serve to smother weeds growing in the crop row  and soil protection (soil building zone).Figure 4). Specifically, we propose that SFZM engenders a self-reinforcing feedback process that couples improvements in soil regulating and supporting services (below-ground cycle) with improvements in provisioning services via increased field working days (above-ground cycle).We base our hypothesis of joint enhancement in provisioning and other ecosystem services via SFZM on a virtuous cycle model that links above-ground and below-ground processes : that SFZM increases field working days by allowing cultivation and planting to occur earlier in the season compared to when these operations could occur, for example, in an adjacent field managed with no-tillage approaches. SFZM would also likely outperform conventional tillage in terms of field working days in poorly drained soils or in years with wet springs .Existing SFZM systems , which remove crop residues from crop row positions prior to planting, have been demonstrated to produce seedbed environments that warm and dry rapidly in early spring . These sFigure 4, points 1 and 2).Soil functional zone management can also extend the growing season by continuing to support crop growth later in the season. Existing SFZM systems concentrate soil moisture into crop inter-row positions , substanFigure 4, point 3). The conservation of soil moisture through late summer in SFZM would also provide a water resource for the establishment of winter crops in double cropping systems, which are currently hampered by growing season duration. By extending the growing season, SFZM has the potential to reduce risks of seasonal crop yield reductions due to delayed harvest under temporal intensification. In addition, the ability of SFZM to enhance soil water conservation could potentially reduce requirements for additional irrigation, as required in some temporally intensified systems .Temporal intensification may itself also help agriculture become more resilient to climate change. For example, double cropping, facilitated by SFZM, may shift phenologies of some crops, enabling them to avoid peak summer temperatures during critical development phases, when excessive heat can cause severe yield reductions . MoreoveFigure 4, point 4). At the most basic level, the production of microbial biomass is governed largely by input quality and microbial physiological traits, such as microbial C-use efficiency .By enabling an extension to the period of living plant cover, SFZM can also promote increases in the production of root exudates and crop residues .Traditional soil models suggest that it is not possible to maintain soil quality under conditions of intensifying production and greater extraction of soil resources, because removal of crop residues and intensification of tillage and fertilization will deplete SOM . This maFigure 4, point 7). In fact, microbial necromass, metabolites, and decomposition products account for the majority of stabilized SOM . By encouraging the development of greater microbial biomass, SFZM may halt declines of SOM observed under conventional tillage, and instead contribute positively to SOM accumulation and soil structure development while simultaneously supporting greater yield extraction through temporal intensification.Although microbial biomass can be rapidly mineralized by soil organisms due to its favorable energy yield and low C:N ratio , microbiized SOM . The accDevelopment and implementation of novel agroecological management systems that allow increases in provisioning services (yield) while simultaneously enhancing regulating and supporting ecosystem services are urgently needed. As our review shows, SFZM offers a strategy for integrating the production benefits associated with intensively tilled field crop production systems with the soil ecosystem service benefits associated with no-tillage. In short, SFZM offers the potential to achieve the best of both approaches. The soil heterogeneity produced by SFZM enhances soil functional biodiversity, and allows farmers to harness this biodiversity to elicit desirable ecosystem functions at appropriate times and places. This can lead to greater resource-use efficiency and closer synchrony between soil processes and crop physiological demands. Moreover, the ability of SFZM to favorably alter soil hydrothermal properties allows extension of the growing season, both at the beginning and end. This opens opportunities for increasing agricultural production via temporal intensification. Coupled with improvements to soil regulating and supporting services, SFZM therefore offers a vehicle for optimizing multiple ecosystem goods and services in agricultural systems.Widespread adoption and refinement of SFZM depends on progress on several fronts. Further research on all aspects of SFZM systems will be required to ensure that service delivery can be optimized to meet specific needs of farmers and society in particular cropping systems and geographies. As well, progress on adoption and refinement of SFZM systems is likely to be strongly affected by societal demand for the full range of regulating and supporting ecosystem services that such systems may be able to provide . The casFigure 1; AG and RS developed Figure All authors contributed to the manuscript by reviewing literature, discussing and developing ideas, writing text sections and revising drafts of the manuscript. AW, DK, AD, AG, SH, MH, RK, DM, RS, SS, KS, AY, and NJ developed the initial idea for the manuscript and all contributed to framing and general writing . PE, LA, and AY contributed to SFZM and Regulating soil services. AJ, ML, and YL contributed to Supporting soil services. AW, AD, AG, RS, and NJ contributed to Virtuous cycles. AW and PE developed The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Aerobic fitness (AF) and self-reported physical activity (srPA) do not represent the same construct. However, many exercise and brain aging studies interchangeably use AF and srPA measures, which may be problematic with regards to how these metrics are associated with brain outcomes, such as morphology. If AF and PA measures captured the same phenomena, regional brain volumes associated with these measures should directly overlap. This study employed the general linear model to examine the differential association between objectively-measured AF (treadmill assessment) and srPA (questionnaire) with gray matter density (GMd) in 29 cognitively unimpaired community-dwelling older adults using voxel based morphometry. The results show significant regional variance in terms of GMd when comparing AF and srPA as predictors. Higher AF was associated with greater GMd in the cerebellum only, while srPA displayed positive associations with GMd in occipito-temporal, left perisylvian, and frontal regions after correcting for age. Importantly, only AF level, and not srPA, modified the relationship between age and GMd, such that higher levels of AF were associated with increased GMd in older age, while decreased GMd was seen in those with lower AF as a function of age. These results support existing literature suggesting that both AF and PA exert beneficial effects on GMd, but only AF served as a buffer against age-related GMd loss. Furthermore, these results highlight the need for use of objective PA measurement and comparability of tools across studies, since results vary dependent upon the measures used and whether these are objective or subjective in nature. A strong link has been established between higher physical activity (PA) levels and improved cognitive function in aging. Epidemiological studies have found that higher midlife PA is associated with lower risk of cognitive decline and dementia later in life , which is usually obtained through a graded treadmill test. The majority of older persons (>75%), however, are unable to satisfactory complete a maximal graded exercise test , which are demonstrated to correlate with VO2max  and PA are used interchangeably; however, they represent different concepts that are measured in specific ways. Using these metrics interchangeably may be problematic with regards to how they are associated with morphological presentation of the brain. If, for example PA and AF measures indexed the same construct, we would expect no difference in neuroimaging assessments when using these measures interchangeably. Simply put, regional brain density would be predicted to be similar when comparing AF and PA measures. To attain an operational definition that could be referenced across studies, Caspersen and colleagues defined these frequently used terms as follows: PA is “any bodily movement produced by skeletal muscles that results in energy expenditure”, while physical fitness is a “set of attributes that people have or achieve and are either health- or skill-related”  have had somewhat overlapping results  and a self-report PA questionnaire (modified version of the Leisure Time Exercise Questionnaire) with GMd in cognitively normal community-dwelling older adults. We hypothesized that GMd associations would differ respective of AF level as compared to self-reported PA (srPA) given the nature of the different constructs and the objective and subjective nature of the indices, respectively. We also investigated whether AF or srPA modified the relationship between GMd and age and hypothesized that AF rather than srPA would modify this relationship given the objective, and thus more precise nature of this measure compared to srPA.Thirty cognitively healthy community-dwelling older adults participated in a study of aging and AF, srPA, functional MRI (fMRI), and structural MRI. fMRI findings for category fluency and finger tapping tasks related to aging and srPA have been reported elsewhere  was used, which is a 3-item scale that asks participants to rate how often they engaged in mild, moderate, and strenuous leisure-time exercise in a certain period of time , with an 8-channel SENSE radio frequency head coil, at the McKnight Brain Institute of the University of Florida. Structural TFE T1-weighted images were acquired for 160 × 1 mm sagittal slices .An MRI screen was conducted via telephone to identify potential study candidates. Those who were MRI-eligible were asked to obtain written clearance from their primary care physician in order to participate in the treadmill test. The 12 min treadmill test took place during the 1st study session during which neuropsychological assessment was also conducted. Heart rate was monitored throughout the treadmill assessment. Prior to commencing the test, participants were asked to take some time to familiarize themselves with the treadmill and the test began once they reported verbally to the study staff that they were comfortable. After the treadmill test, participants were instructed on how to answer the modified LTEQ, which they completed daily for 7 days following the 1st study session. Structural brain imaging scans were obtained during the 2nd session following the 7 day srPA monitoring period.MRI structural images were analyzed using FSL-VBM, a voxel-based morphometry (VBM) analysis  using the Threshold-Free Cluster Enhancement (TFCE) method. This method enhances cluster-like regions more than background (noise) by creating output that is a weighted sum of all of the local clustered signal, without the need for arbitrary cluster thresholding. If the voxel-wise analyses returned any significant interaction terms (age*AF or age*srPA), the average per-subject GMd values were extracted from significant clusters and further analyzed using IBM SPSS (version 21) to characterize the direction of the interaction.We used the randomize option in FSL, which is a permutation-based program allowing for modeling and inference testing using standard general linear model design, when the null distribution of a statistic map is unknown. To describe the unique relationship between GMd, age, fitness level, and srPA, we conducted two voxel-wise multiple regression models under the general linear model, higher level/non-time series design option in FSL: (1) The first regression model used voxel-wise GMd as the dependent variable and age, AF (distance traveled in 12 min during treadmill test), and the interaction term between age and AF as independent variables; and (2) The second regression model included voxel-wise GMd as the dependent variable and age, srPA (self-reported minutes of moderate + strenuous activity in 7 days), and the interaction term between age and srPA as independent variables. These two whole-brain analyses were conducted using data from all 29 participants, and the independent variables were centered (demeaned) and continuous. The resulting probability maps were corrected for family-wise error (post hoc region of interest (ROI) analysis was conducted using anatomically-defined ROIs based on the cortical and subcortical Harvard-Oxford atlases available in FSL. Pearson bivariate correlations were conducted between mean GMd values extracted from each ROI and AF and srPA using IBM SPSS. ROIs included: right and left hippocampi, inferior and middle frontal gyri, frontal pole, anterior cingulate cortex, and the angular and supramarginal gyri.Furthermore, given the reported associations between GMd and the hippocampus, anterior cingulate cortex, lateral frontal, and lateral parietal regions  bilateral occipital poles, lingual and left fusiform gyri, and right calcarine cortex . The interaction term between age and AF was significantly associated with GMd in three clusters: (1) bilateral cerebellum, lingual, and fusiform gyri, and left inferior and middle temporal gyri , and the interaction term between age and AF as independent variables. It is important to note that AF was dichotomized into high and low fitness groups using a median split in order to characterize the interaction term and not for the purposes of comparing low and high AF groups. For those with lower AF, there were significant negative associations between age and GMd in these three regions, while for individuals with higher fitness, there was a significant positive association between age and GMd in temporo-cerebellar regions. See Table r = −0.403, p = 0.03, N = 29).There were no significant associations between the AF measure and GMd in any of the ROIs . srPA was associated with GMd in the left hippocampus only , they represent different constructs. Hence, it is not surprising that they predict differential brain morphology when the variance for age within this older sample is removed. The second factor is that our measure of AF was objective and our measure of PA was self-report. The lower reliability  of self-report measures may account for the lack modulation of age-related loss in GMd by PA. It is critical to know whether PA can modulate age-related GMd loss. Hence, future studies investigating these associations should rely on objective measurement of PA since srPA is frequently overestimated . Similarly, joint pain, which is common in the elderly and affects the ability to stand or walk (Keenan et al., The present study is limited by a small sample size and by the fact that the In conclusion, the current study found no overlap in regional GMd associated with objectively-measured AF and a self-report measure of PA in cognitively unimpaired older adults, highlighting the need to better understand the associations between brain outcomes and different measures of AF and PA. We also found that the relationship between GMd and age changed as a function of AF, but not srPA, attesting to the neuroprotective role of AF in aging by sparing GMd loss as age increases. Future studies investigating the underlying mechanisms by which exercise improves brain health should use objective measurement of PA and AF that is comparable across studies.All authors contributed substantially with conception and study design, data acquisition and analyses, data interpretation, manuscript drafting and revisions, and manuscript approval.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Ventricular noncompaction (VNC) of the myocardium is a rare genetic cardiomyopathy caused by a disorder during endocardial morphogenesis and could be accompanied by life-threatening complications. The major clinical manifestations of VNC are heart failure, arrhythmias, and embolic events. The left ventricle is the most commonly reported affected site, but a few cases of right ventricular involvement have also been reported. We report a case of biventricular noncompaction cardiomyopathy in a 31-year-old woman presenting with a new onset seizure. On the second day of her telemetry-monitored hospitalization, she suffered a witnessed ventricular fibrillation arrest requiring emergency direct-current cardioversion and induced hypothermia. Transthoracic echocardiography (TTE) showed isolated left ventricular (LV) noncompaction and depressed LV systolic function. Subsequent cardiac magnetic resonance imaging (MRI) revealed both left and right ventricular noncompaction. This unusual presentation highlights the importance of a complete and thorough evaluation of patients even when presenting with apparently noncardiac symptom(s). Ventricular noncompaction (VNC) of the myocardium is a rare genetic cardiomyopathy that is believed to arise from arrested endomyocardial development during embryogenesis . It is aA 31-year-old woman presented with a witnessed loss of consciousness while driving. The episode lasted for about five-to-ten minutes and was associated with tonic-clonic seizure-like movement. There was no urinary or bowel incontinence. She had no history of seizures, arrhythmias, or family history of sudden cardiac death.Physical exam was remarkable only for a 2/6 systolic ejection murmur loudest at the left midsternal border on cardiac auscultation. The laboratory results were within normal limits. Electrocardiogram (ECG) showed sinus rhythm with occasional premature ventricular complexes. An electroencephalogram (EEG) and a head-computed tomography (CT) scan were unrevealing.In the course of the workup, the patient suffered a ventricular fibrillation cardiac arrest requiring defibrillation and induced hypothermia for resuscitation. She recovered without neurologic deficits. Cardiac evaluation with echocardiogram  showed dVNC of the myocardium is a rare genetic cardiomyopathy. The left ventricle is more frequently involved, but biventricular involvement, as in this case, is rarely encountered , 11. WhaAlthough tonic-clonic seizure-like activity was the only presenting symptom given the extensive structural heart disease, it is plausible that this patient likely had a ventricular arrhythmic event  leading to generalized cerebral hypoxia, and thus the tonic-clonic seizure activity. The resulting motor activity has been attributed to a decreased cerebral blood flow , which rAlthough left ventricular noncompaction is more common, the improved imaging capabilities of cardiac MRI provide opportunity for better characterization of right ventricular involvement. Echocardiography has a crucial role in the diagnosis of VNC, but cardiac MRI, contrast ventriculography, and computed tomography could also be utilized during assessment, especially in patients with poor image quality on echocardiography , 22, in In conclusion, VNC is an uncommon disorder accompanied by life-threatening complications. Its unusual presentation highlights the importance of a complete and thorough workup of patients presenting with apparently noncardiac symptom(s). Data is limited on specific therapy for VNC, but it is recommended that medical management be tailored towards the clinical manifestations, and standard guidelines should be applied for patients with reduced LVEF, and heart failure with preserved systolic function , in the


Retinoic acid is a derivative of vitamin A and is involved in the proliferation and differentiation of stem/progenitor cells in several tissues. However, whether retinoic acid regulates the characteristics of HpSCs in the normal liver is still unknown. A recent study has shown that acyclic retinoid regulates the survival and proliferation of HpSCs derived from mouse foetal liver. Acyclic retinoid suppressed the expansion of CD29 Stem Cell Research & Therapy, investigated a new mechanism regulating the survival and differentiation of these stem/progenitor cells by retinoic acid signals [The liver is the largest organ in the adult human body and is important for maintaining homeostasis. Adult hepatocytes and liver parenchymal cells express genes and enzymes for several metabolic functions. In addition, the liver includes several non-parenchymal cells such as cholangiocytes, sinusoidal endothelial cells, stellate cells, and Kupffer cells. Hepatocytes and cholangiocytes are differentiated from the same source: the stem/progenitor cells in embryonic livers. Guan and colleagues, in a recent article in  signals .There are two types of liver progenitor cells: foetal hepatic progenitor cells and postnatal/adult hepatic progenitor cells. Liver formation begins with the specification of the foetal hepatic progenitor cells derived from the foregut endoderm. During mid- to late-foetal liver development, hepatic progenitor cells differentiate into hepatocytes and cholangiocytes. It has been reported that the interaction between hepatic cells and hematopoietic cells is important for foetal liver maturation . Moreovein vitro colony formation assays of hepatic progenitor cells. In addition, markers such as Dlk, Liv2, E-cadherin, CD13, and CD133 are reported to be specific markers of foetal hepatic progenitor cells. CD13 and CD133 are also expressed in progenitor cells derived from adult livers. Recent studies revealed that long-term proliferative hepatic progenitor cells exist in mouse and human adult livers. LGR5+ and EpCAM+ progenitor cells can form cystic structures and expand in vitro for several months [in vitro culture system of purified cells using antibodies against these cell surface antigens is useful for analyses of molecular mechanisms regulating the proliferation and differentiation of hepatic progenitor cells.These foetal and adult hepatic progenitor cells are well characterized by specific cell surface antibodies and cell purification methods such as flow cytometry. Suzuki and colleagues  found CDl months . An in vin vivo is induced by the addition of all-trans retinoic acid (ATRA) [It is known that, in addition to stem/progenitor cells in the normal liver, cancer stem-like cells are present in hepatocellular carcinomas (HCCs) or other types of cancers. Cancer stem-like cells have characteristics similar to those exhibited by somatic stem cells and are important for tumour initiation, metastasis, and recurrence. Retinoic acid is a natural derivative of vitamin A and regulates its target genes by binding to nuclear receptors such as retinoic acid receptors (RARs) and retinoid X receptors (RXRs). Retinoic acid is important for the differentiation and proliferation of stem/progenitor cells in both somatic and cancer tissues. For example, retinoic acid is used for the differentiation of pluripotent stem cells into mature functional cells. In addition, acyclic retinoid (ACR) exhibited anti-HCC effects in several hepatoma models , 8. In cd (ATRA) . Thus, tCip1 was upregulated by the addition of peretinoin. These results suggested that ACR inhibits the expansion of stem/progenitor cells derived from embryonic livers. This cell cycle arrest is involved in the differentiation of HpSCs. The authors found that ACR induced maturation of HpSCs into hepatocytic cells. The addition of peretinoin induced the expression of cells positive for the hepatocyte marker albumin in this culture system. In contrast, the HpSC marker genes such as α-feto protein, Cd44, and Dlk were significantly downregulated by peretinoin. The expression of cytokeratin 19, a cholangiocyte marker, was also decreased, suggesting that the addition of ACR promoted hepatic differentiation, but not cholangiocytic differentiation, of HpSCs. In addition, ACR regulated HpSC survival via apoptosis.Guan and colleagues  stated tGuan and colleagues have provided evidence that retinoic acids regulate proliferation and differentiation of HpSCs during liver development. Whether retinoic acids ATRA and ACR positively or negatively affect cell proliferation in normal liver and hepatic carcinoma tissues is controversial. The present study clearly showed that ACR suppressed the proliferation and survival of stem/progenitor cells in normal embryonic liver tissues. A synthetic retinoid ACR might have a cell signal cascade that is different from that of natural retinoic acids. Hepatic cancer stem cells also exhibit stem/progenitor characteristics similar to those exhibited by normal HpSCs. Therefore, this study suggested that ACR not only plays the role of an anti-tumour drug in HCC but also inhibits tumour-initiating cells such as hepatic cancer stem cells.


Atomistic molecular dynamics simulations were performed with 13 non-peptidic neurotransmitters (NTs) in three different membrane environments. The results provide compelling evidence that NTs are divided into membrane-binding and membrane-nonbinding molecules. NTs adhere to the postsynaptic membrane surface whenever the ligand-binding sites of their synaptic receptors are buried in the lipid bilayer. In contrast, NTs that have extracellular ligand-binding sites do not have a similar tendency to adhere to the membrane surface. This finding is a seemingly simple yet important addition to the paradigm of neurotransmission, essentially dividing it into membrane-independent and membrane-dependent mechanisms. Moreover, the simulations also indicate that the lipid composition especially in terms of charged lipids can affect the membrane partitioning of NTs. The revised paradigm, highlighting the importance of cell membrane and specific lipids for neurotransmission, should to be of interest to neuroscientists, drug industry and the general public alike. Neurotransmission regulates a variety of senses and functions such as motivation, memory and muscle contraction. Neurotransmitters (NTs) play a key role in these functions since the signal transmission takes place through the binding of a NT with its specific receptor embedded in the postsynaptic membrane. Understanding the details of NT binding is important given that various diseases emerge if the signaling is impaired.Currently, NTs are depicted in neurobiology textbooks to travel across the synaptic cleft over a distance of 20–40 nm and to bind to their receptors without interacting with the membrane itself1bc1 complex operation9101112910Until a few decades ago, membranes were considered as passive participants in cellular processes. Recently, however, lipids and membranes have been shown to be in a central position in numerous cellular functions such as signaling and protein sorting457The effect of lipids and cell membranes is not limited to proteins and peptides but the dynamics of ligands and other small molecules is affected, too. For instance, antipsychotic drugs have been shown via numerous biophysical methods to interact strongly with lipid membranes131518193 muscarinic acetylcholine receptor, the lipid composition is a factor that should not be overlookedA substantial body of work indicates that some ligands first bind to the membrane surface and then diffuse laterally into the membrane-buried binding sites of their receptors2223242526Several experimental and computational studies have indicated that the lipid composition in membranes has a role in NT dynamics. The lipid composition has been shown by molecular dynamics (MD) simulations and calorimetry29313435In this work we performed atomistic MD simulations for three lipid bilayer models with 13 non-peptidic NTs and the dopamine precursor L-dopa. The pre-equilibrated lipid membrane models Based on the MD simulation results the current view of neurotransmission, which does not account for the role of lipids1As expected, the NTs did not permeate the lipid bilayers; instead they displayed varying degrees of reversible membrane attachment. Catecholamines such as dopamine and norepinephrine, as well as serotonin, melatonin, and adenosine, all attached strongly to the lipid bilayers in the NT-membrane simulations ; Table 1The simulations indicate that NTs can be divided into membrane-binding  and membBased on the divergent behavior observed in the simulations, neurotransmission can be split into two different mechanisms . The claAlthough the lipid types residing in synapses are known, the exact lipid compositions of specific synapses or, more importantly, their extracellular and intracellular leaflets are yet to be characterized394041The ligand-binding sites of G protein-coupled receptors are typically membrane-buried  and theiThe ligand-gated ion channels such as ionotropic glutamate receptors have extracellular ligand-binding sites. Accordingly, their representative NTs follow the membrane-independent mechanism . GABA, gThe above described results suggest that specific lipid types and the lipid composition in general could have an important role in synaptic neurotransmission. While the concentrations of negatively charged glycolipids are usually small in typical animal cells, their levels are known to be higher in neurons39Tightly regulated lipid composition differences between synapses could ensure that acetylcholine can bind into both membrane-buried and extracellular ligand-binding sites . In fact434445Synaptic receptors are not the only membrane-embedded proteins that interact with NTs at the postsynaptic membrane. The timely removal of NTs from the synaptic cleft is vital and, for example, if glutamate transport is obstructed, neurotoxic effects follow+ gradient across the membrane powers the Na+/NT co-transport. In addition, the positive ion(s) at the ligand-binding sites form ionic bonds with NTs that have carboxylate groups  simulations were performed with GROMACS 4.53852How to cite this article: Postila, P. A. et al. Selective effect of cell membrane on synaptic neurotransmission. Sci. Rep.6, 19345; doi: 10.1038/srep19345 (2016).