Specificity of ioflupane [123I] imaging for bradykinetic rigid or posturally (BRP) unstable motor subtype was lower, but better than for baseline clinical diagnosis.Diagnostic effectiveness of ioflupane [123I] imaging for clinical diagnosis of Parkinson’s syndrome (PS) or non-PS was associated with tremor and balanced motor dominance rather than with BRP dominance. High diagnostic effectiveness of ioflupane [123I] imaging and favourable performance relative to final clinical diagnosis at 1 year post-scan in subjects with CUPS was demonstrated. This study suggests that the diagnostic performance of ioflupane [123I] imaging in CUPS remains high at all stages of disease, including early stage, and across both age groups and cognitive state (MMSE).Strongest diagnostic performance of ioflupane [ We report the diagnostic effectiveness , negative predictive value (NPV), and accuracy) of ioflupane [123I] imaging versus the final clinical diagnosis for subjects who had both ioflupane [123I] imaging results at baseline and a specific clinical diagnosis 1 year post scan, including analyses based on subject’s Hoehn and Yahr (H&Y) stage, Mini-Mental State Examination (MMSE) score, age, and predominant clinical syndrome subgroup. It has already been established that age is a contributing factor in the progressive decline of dopamine transporter (DaT) binding in healthy aging subjects  imaging results. and 1-year clinical diagnosis) were included in this report. To analyze the effect of the H&Y stage on diagnostic effectiveness of ioflupane [123I] imaging, subjects were classified using a baseline-modified H&Y stage dichotomized into two groups: stage <2 and stage ≥2. To analyze the effect of cognitive status of subjects on diagnostic effectiveness of ioflupane [123I] imaging, subjects were grouped by scores <29 and ≥29 . To analyze the effect of subject’s age on the diagnostic effectiveness of ioflupane [123I] imaging, subjects were classified as <68 or ≥68 years of age at baseline. Investigators also grouped subjects at baseline into a predominant clinical motor syndrome subgroup – tremor dominant; bradykinesia, rigidity, postural instability (BRP) dominant; or balanced (regarding tremor and bradykinesia scores) – to determine the effect of motor symptom subtype on diagnostic effectiveness of ioflupane [123I] imaging.The clinical and imaging methods were reported previously . Ioflupa the USA ,7]. Of . Of 123ISubjects in the tremor dominant subgroup had tremor scores greater than their maximal score for bradykinesia, rigidity, or postural instability. Subjects in the BRP dominant subgroup had the maximal score of bradykinesia, rigidity, or postural instability greater than their score for tremor. Subjects in the balanced group had maximal BRP scores equal to tremor scores. The scores were calculated by assigning numbers scores for motor signs: 1 for none, 2 for possible, and 3 for definite.123I] imaging results in subjects with CUPS [123I] imaging, which in combination with 1-year post-scan clinical follow-up provided sufficient rationale for using the 1-year clinical diagnosis as a reference standard (standard of truth). The study population receiving ioflupane [123I] imaging from which the 92 subjects were selected to be analyzed had been symptomatic for 2.54 years (mean) or 2.20 years (median)  SPECT imaging methodology has been reported previously [123I] SPECT image was categorized as normal or abnormal by an onsite imaging reader (a nuclear medicine physician having expertise in neuroimaging) and interpreted with no access to clinical information, such as symptoms, clinical signs evolution, treatment, and clinical management changes.Description of the ioflupane 23I SPECT123I] imaging in subject subgroups. The subgroups for H&Y stage, MMSE, and subject’s age were based on dichotomization; that is, an entire population at the median values (to ensure the same number of subjects in low and high subgroups). Dichotomization is a common statistical approach used with limited datasets for which categorical, more meaningful subdivisions requiring a much larger sample size cannot be used. The median values were chosen as the cutoff points to avoid subjectivity by the authors and to prevent outliers from having undue influence. The values we used were H&Y median score of 2, MMSE median score of 29, and median age of 68 years, respectively. The subgroups for dominant motor syndrome were based on a clinically common classification into tremor dominant, BRP dominant, and balanced subgroups. Standard methods were employed to calculate the diagnostic efficacy parameters for ioflupane [123I] imaging using 1-year post-scan clinical diagnosis as the reference standard (standard of truth). For subjects with a PS diagnosis at 1 year, an ioflupane [123I] image reading of abnormal was deemed a true positive, whilst an ioflupane [123I] reading of normal was deemed a false negative. For subjects with a non-PS diagnosis at 1 year, an ioflupane [123I] image reading of normal was deemed a true negative, whereas an ioflupane [123I] image reading of abnormal was deemed a false positive. Comparison of diagnostic efficacy parameters between mutually exclusive subgroups was performed with Fisher’s exact test.The statistical analysis plan was to generate diagnostic efficacy parameters – sensitivity , specificity , PPV, NPV, and accuracy – for ioflupane [123I] imaging results among all subgroups. Table 123I] imaging results.The number of subjects analyzed in four subgroups was 92, with approximately equal distribution between normal and abnormal ioflupane 23I imagi123I] images. The subjects who had abnormal ioflupane [123I] images were about evenly split between H&Y stages 1 and 2.At baseline, most of the 92 subjects in this group were in the early stages of disease (H&Y stages 1 to 2), split about evenly between the two stages. Slightly less than one-half of the subjects had normal ioflupane [123I] imaging ranged between 91 and 100% to determine whether or not subjects had PS or non-PS at 1 year for all diagnostic efficacy parameters: sensitivity, specificity, accuracy, PPV, and NPV. In addition, all of the efficacy parameters showed similar high values for subject groups having either low (<2) or high (≥2) H&Y stage scores  in both groups. Overall, no noticeable difference was observed between age groups for all tested parameters.At baseline, subjects were divided relatively equally between the younger group (<68 years of age) and the older group (≥68 years of age) .Subjects with baseline motor signs with predominance of bradykinesia, rigidity, or postural instability had moderate (ranged between 67 and 89%) sensitivity, accuracy, and PPV of ioflupane 23I imagi123I] imaging in subjects with tremor dominant and balanced motor signs ranged between 92 and 100%, showing specificity of 100% for the tremor dominant and balanced motor signs subgroups  found significant differences for the tremor dominant and balanced subgroups  as compared with the baseline clinical diagnosis (ranged between 44 and 66%). For the balanced subgroup, specificity, PPV, and accuracy were significantly higher for ioflupane [123I] imaging  as compared with the baseline clinical diagnosis .Comparison of the specificity, PPV, and accuracy of ioflupane 13I imagin123I] imaging as an adjunct to clinical diagnosis. This study is the first robust statistical analysis of several potential influences on the efficacy of ioflupane [123I] imaging that measures their effect on image interpretation and supports the value of imaging in a real-world setting where clinicians are able to incorporate imaging results to assist them in making a diagnosis and subsequent treatment plan. The open-label setting is both a strength and a possible limitation of the study. Whilst the findings of the study indicate some utility of ioflupane [123I] in assisting diagnosis, one of the possible methodological limitations of this study is that information from the imaging scan could have influenced the clinician’s standard-of-truth diagnosis, causing a shift in the subsequent clinical management of the subject [123I] imaging between different motor syndrome subgroups, including subjects with CUPS. Because of the modest study size and possible unequal distribution of subjects amongst motor syndromes, the small number of participants in the BRP dominant subgroup may not be sufficiently powered to adequately address real differences in imaging utility from other motor subgroups. Despite this, the overall accuracy of ioflupane [123I] imaging for the BRP dominant subgroup was still very good, although less than for the tremor dominant and balanced subgroups.The findings in this study are broadly in line with hypothetical predictive models and augment the scant literature examining the effects of age, clinical stage of PD and cognitive impairment on the utility of ioflupane 23I imagi123I] imaging abnormality in patients at initial motor presentation of PD in a UK movement disorder service noted the majority of patients to be at stage 2 to 3 of ioflupane [123I] imaging abnormality [123I] imaging might be able to detect abnormality both at early (premotor) and later (motor) clinical presentation, although to date there has been no analysis of H&Y stage versus clinical utility of ioflupane [123I]. The findings in our study support this hypothetical notion of no significant effect of H&Y stage on diagnostic accuracy of ioflupane [123I].It has been established that SPECT imaging of presynaptic DaTs has utility in the premotor diagnosis of PD, as noted in analysis of cohorts presenting with REM sleep behavioral disorder or anosomia/hyposmia ,9]. Fur. Fur9]. ormality ,11]. Ta. Ta123I]123I] imaging varies with stage of cognitive impairment in CUPS has also not been analyzed to date. The findings in our study of no effect of MMSE on accuracy of ioflupane [123I] imaging suggest the technique remains accurate in the context of cognitive impairment. Our analyses did not include subgrouping of mild, moderate, or severe dementia based on MMSE, which would have been useful, although data from the DLB Study Group would suggest the technique could be of diagnostic utility even in the presence of severe cognitive impairment [Whether or not diagnostic utility of ioflupane 3I imaginpairment .123I] imaging was to show subtle or minimal change between normal and abnormal cases. Previous reports correlating ioflupane [123I] image abnormality with first clinical presentation of PD [123I] image abnormality at motor PD presentation is in line with an estimated 60 to 80% dopaminergic cell loss at motor presentation, and is in any case associated with asymmetrical striatal signal loss rather than the symmetrical loss due to ageing. In accord with the published literature, this would be expected to yield substantial ioflupane [123I] imaging abnormality, and the majority of patients in this study had grade 2 to 3 abnormality (moderate–severe) rather than grade 1 (mild). The mild age-related symmetric reduction in DaT binding with increased age would therefore probably not have a significant effect on diagnostic performance of this test, and the findings in our study are in keeping with this hypothesis.An average age-related decline of DaT availability of 5.5% across both genders per decade has been reported from work on normal healthy controls . This reon of PD ,11], ho, ho123I]on of PD  suggesti123I] image abnormality seen in the motor subtype of PD analyzed, broadly tremor dominant versus akinetic-rigid phenotypes [123I] imaging in making an accurate clinical diagnosis in the two phenotypes, although the areas of reduced dopaminergic projection differ between the two, with visible reduction to the dorsal putamen in akinetic-rigid patients and in the caudate nucleus in tremor dominant patients [123I] imaging scans [123I] imaging for the tremor dominant and balanced subgroups was significantly better than for the baseline clinical diagnosis; for the BRP dominant subgroup, the difference did not reach statistical significance. The specificity of baseline clinical diagnosis for BRP was inferior to ioflupane [123I] imaging performance, supporting challenges for differential work-up in this motor syndrome, especially in subjects with CUPS. One strategy for improving the performance of the ioflupane [123I] imaging in the BRP subgroup could be a quantitative evaluation of SPECT images. Additionally, repeated ioflupane [123I] imaging scanning  could be helpful in those BRP patients with initial normal scans to determine progression of the disease.There have been a number of reports on the pattern of ioflupane . Th. Th123I]ng scans . Notablyng scans . Conceiv123I] imaging, with tremor dominant and balanced motor scores being more highly predictive of PS than bradykinesia, rigidity, or postural instability dominance. In our trial, the type of dominant motor signs, but not the H&Y stage, MMSE score, or age, impacted the diagnostic performance of ioflupane [123I] imaging. Ioflupane [123I] imaging had the strongest efficacy in tremor dominant and balanced cases of CUPS.According to our findings, the type of initial motor signs has an observable relationship with the predictability of performance of ioflupane [123I] imaging in CUPS remains high at all stages of disease, including early stage, and across both age groups and cognitive state (MMSE). The strongest diagnostic performance of ioflupane [123I] imaging for clinical diagnosis of PS or non-PS was associated with subjects with tremor dominant and balanced motor scores rather than with BRP dominance. High diagnostic effectiveness of ioflupane [123I] imaging in diagnosis of CUPS was demonstrated by this study. Favorable performance of ioflupane [123I] imaging was observed relative to final clinical diagnosis at 1 year post scan.This study suggests that the diagnostic performance of ioflupane [BRP: bradykinesia, rigidity, postural instabilityCUPS: clinically uncertain parkinsonian syndromeDaT: dopamine transporterH&Y: Hoehn and YahrMMSE: Mini-Mental State ExaminationNPV: negative predictive valuePD: Parkinson’s diseasePPV: positive predictive valuePS: parkinsonian syndromeSPECT: single-photon emission computed tomographyCC is an employee of H2O Clinical LLC. IDG was an employee of GE Healthcare during the conduct of the study. The remaining authors declare that they have no competing interests.NB participated in the research project  and in manuscript preparation (review and critique). RAH participated in the research project  and in manuscript preparation (review and critique). JS participated in the research project  and in manuscript preparation (review and critique). AK participated in the research project  and in manuscript preparation (review and critique). MP participated in the research project  and in manuscript preparation (review and critique). CC was responsible for the statistical analysis  and participated in manuscript preparation (review and critique). CC takes responsibility for the integrity of the data and the accuracy of the data analysis. IDG participated in the research project  and in manuscript preparation . IDG was a Global Medical Director of the Phase 4 Clinical Trial. All authors read and approved the final manuscript.Presenting the independent ethics committees and institutional review boards approving the clinical trial.Click here for file


Ovarian and triple-negative breast cancers with BRCA1 or BRCA2 loss are highly sensitive to treatment with PARP inhibitors and platinum-based cytotoxic agents and show an accumulation of genomic scars in the form of gross DNA copy number aberrations. Cancers without BRCA1 or BRCA2 loss but with accumulation of similar genomic scars also show increased sensitivity to platinum-based chemotherapy. Therefore, reliable biomarkers to identify DNA repair-deficient cancers prior to treatment may be useful for directing patients to platinum chemotherapy and possibly PARP inhibitors. Recently, three SNP array-based signatures of chromosomal instability were published that each quantitate a distinct type of genomic scar considered likely to be caused by improper DNA repair. They measure telomeric allelic imbalance (named NtAI), large scale transition (named LST), and loss of heterozygosity (named HRD-LOH), and it is suggested that these signatures may act as biomarkers for the state of DNA repair deficiency in a given cancer.We explored the pan-cancer distribution of scores of the three signatures utilizing a panel of 5371 tumors representing 15 cancer types from The Cancer Genome Atlas, and found a good correlation between scores of the three signatures (Spearman’s ρ 0.73–0.87). In addition we found that cancer types ordinarily receiving platinum as standard of care have higher median scores of all three signatures. Interestingly, we also found that smaller subpopulations of high-scoring tumors exist in most cancer types, including those for which platinum chemotherapy is not standard therapy.Within several cancer types that are not ordinarily treated with platinum chemotherapy, we identified tumors with high levels of the three genomic biomarkers. These tumors represent identifiable subtypes of patients which may be strong candidates for clinical trials with PARP inhibitors or platinum-based chemotherapeutic regimens.The online version of this article (doi:10.1186/s40364-015-0033-4) contains supplementary material, which is available to authorized users. Personalized medicine in cancer aims to improve treatment outcome by matching the specific biological characteristics of a tumor with the most appropriate therapeutic option. One such biological characteristic is a defect in one of the DNA repair systems, which often leads to the accumulation of genomic damage, such as point mutations and short indels, as well as gross copy number aberrations, termed genomic scars, that may be gain or loss of large chromosomal regions, or even whole chromosomes ,2. The aNumber of telomeric Allelic Imbalances (NtAI), published by us, was based on a type of genomic scar accumulation which could predict response to platinum-based chemotherapy in triple negative breast cancer  and high grade serous ovarian cancer [Large Scale Transition (LST), published by Popova et al., was based on a type of genomic scar which was associated with loss of BRCA1 or BRCA2 in TNBC [Homologous Recombination Deficiency score (HRD-LOHa), published by Abkevich et al., was based on a type of genomic scar that was enriched in high grade serous ovarian cancer patients with loss of BRCA1 or BRCA2 [Three signatures, each measuring a specific type of genomic scar in SNP array data, were published in 2012 by three groups, including ours. The n cancer . Large S in TNBC . The Homor BRCA2 . The genAs these signatures measure similar genomic features and have been designed for the same purpose of guiding precision medicine, we here use The Cancer Genome Atlas (TCGA) to determine the distribution of the three signatures of genomic scars across 15 different cancer types. We find that cases with high levels of each signature can be found in 12 of the 15 cancer types, suggesting that it may be possible to use genomic scar signatures to identify patients with DNA repair deficiency predictive of platinum chemotherapy sensitivity prior to initiating therapy. We also investigate the association between high levels of each signature and increased ploidy, p53 loss, and other measures of genomic instability.We first determined the distribution of the scores of each signature in the 15 cancer types Figure , Table 1We compared the median signature counts across cancer types and observed that the highest scoring cancer type for the NtAI and LST signatures was serous ovarian cancer  between each pair of signature scores across all cancer types and within each cancer type Figure A. We fouAs NtAI, LST and HRD-LOH can be considered a type of genomic instability score, we next compared the signature scores to three other previously published measures of chromosomal instability: the weighted genome integrity index (wGII) , the freBoth the NtAI and LST signatures measure a genomic aberration pattern where a single allelic event may increase the score. As such, an increase in sample ploidy increases the amount of DNA that may experience genomic aberrations, and this may correlate with increased signature counts. Indeed, in the original publication of LST , Popova When we compared the signature scores in near-diploid versus near-tetraploid cancers see , we do iAs both TNBC and serous ovarian cancer commonly have p53 mutations, we tested if p53 status itself would be associated with increased scores of NtAI, LST and HRD-LOH. We examined cancer types with at least 20 samples with P53 loss, and at least 20 samples with wtP53. We found that all showed a significant increase in the scores of all signatures in mP53, with the exception of melanoma and stomach adenocarcinoma for LST, and melanoma for HRD-LOH  versus high stage (stage 3 + 4), we found that high stage was significantly associated with higher NtAI and LST scores in breast and head and neck cancer, and with higher of all three scores in colon, renal, and endometrial cancer , and compared these to patients that never smoked, or stopped smoking at least 15 years ago (“non-smokers”). This showed a clear increase in the median NtAI, LST and HRD-LOH counts for smokers relative to non-smokers in head and neck cancer patients , and in lung adenocarcinoma cancer . No significant difference was found for bladder or squamous cell lung cancer  to measure concordance between continuous signature scores. The Wilcoxon rank sum test was used to test for increase in signature scores by ploidy. All p-values are two-sided.aIn the original publication [lication  this metlication , and sublication  is curre


The intestinal epithelium has a strategic position as a protective physical barrier to luminal microbiota and actively contributes to the mucosal immune system. This barrier is mainly formed by a monolayer of specialized intestinal epithelial cells (IECs) that are crucial in maintaining intestinal homeostasis. Therefore, dysregulation within the epithelial layer can increase intestinal permeability, lead to abnormalities in interactions between IECs and immune cells in underlying lamina propria, and disturb the intestinal immune homeostasis, all of which are linked to the clinical disease course of inflammatory bowel disease (IBD). Understanding the role of the intestinal epithelium in IBD pathogenesis might contribute to an improved knowledge of the inflammatory processes and the identification of potential therapeutic targets. Inflammatory bowel disease (IBD) is characterized by a chronic idiopathic inflammation of the intestine and consists of two main forms, i.e., ulcerative colitis (UC)  and CrohThe intestinal epithelium forms the protective barrier and host defense against the harmful luminal microenvironment with selective permeability and absorption of nutrients. The epithelium is covered by a single-cell layer composed of different subtypes of specialized intestinal epithelial cells (IECs) including absorptive cells, goblet cells, enteroendocrine cells, Paneth cells, M cells, cup cells, and Tuft cells, all of which differentiate from epithelial stem cells –18. ThesThe epithelial monolayer is the main component of the epithelial barrier and its ability to act as a protective physical barrier is mediated by the formation of a web of tight junctions (TJs) that regulate the paracellular permeability and barrier integrity, production of mucus layer covering the luminal surface of the epithelium, and recognition of pathogens and production of antimicrobial peptides (AMPs) to ensure effective immunity . Hence, Muc2-deficient mice -like receptors in the cytoplasm that are essential in sensing bacterial products and initiating the immune response through the pro-inflammatory transcription factor nuclear factor-κB to maintain homeostasis . TherefoGenes within IBD-associated genetic loci highlight the importance of epithelial barrier defects . While tHNF4α, regulates the expression of several components involved in epithelial TJs and intestinal permeability. HNF4α is down-regulated in patients with IBD and, moreover, mice with IEC-specific conditional knockout of Hnf4α are more susceptible to chemically induced colitis , encoded by  colitis , indicatferation . Hence, feration .meprin 1A (MEP1A) gene have been associated with UC  protein, which is encoded by ATG16L1  can be maintained long-term elevance . Within The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In vivo genetic manipulation has implicated a variety of candidate directional signals, cell surface receptors, and signaling pathways, that may be involved in polarizing multipolar cells and stabilizing a pia-directed leading process for radial migration. Other signals are implicated in starting multipolar migration and triggering axon outgrowth. Here we review the molecules and mechanisms that regulate multipolar migration, and also discuss how multipolar migration affects the orderly arrangement of neurons in layers and columns in the developing neocortex.Most neurons migrate with an elongated, “bipolar” morphology, extending a long leading process that explores the environment. However, when immature projection neurons enter the intermediate zone (IZ) of the neocortex they become “multipolar”. Multipolar cells extend and retract cytoplasmic processes in different directions and move erratically—sideways, up and down. Multipolar cells extend axons while they are in the lower half of the IZ. Remarkably, the cells then resume radial migration: they reorient their centrosome and Golgi apparatus towards the pia, transform back to bipolar morphology, and commence locomotion along radial glia (RG) fibers. This reorientation implies the existence of directional signals in the IZ that are ignored during the multipolar stage but sensed after axonogenesis.  The neocortex develops by the coordinated migration of projection neurons from the neocortical ventricular zone (VZ) and interneurons from the ganglionic eminences Hatten, . While pThis review addresses new developments and continuing uncertainty regarding the external signals and intracellular mechanisms that regulate MP cells at three important times: when MP migration starts, when the axon starts to grow, and when MP cells repolarize towards the pia and resume radial migration with bipolar morphology. The review also discusses the implications of MP migration for the radial unit hypothesis of cortical wiring and for cortical lamination. The focus is on newer research from the last 5–10 years, with an emphasis on loss of function studies. Readers are referred to excellent reviews for earlier work   member that is not secreted, but that acts inside the cell through mechanisms that are receptor-independent. Wu et al showed that FGF13 localizes in growth cones of cultured neurons, binds to microtubules, and stimulates microtubule polymerization , allowing separation of apical junctional complexes : p27 expression is inhibited by Cx43 knockdown and increased by Cx43 over-expression  , which is absent from thalamocortical axons in the upper IZ  moiety and is localized to lipid rafts, where it potentially activates Src-family kinases. Indeed, Namba et al show that the Src family kinase (SFK) Lyn and small GTPase Rac1 are likely involved in stabilizing axons downstream of TAG1  and TrkC (Ntrk3) and ligands BDNF and NT3 are expressed in the VZ/SVZ. Trk inhibitors inhibit axonogenesis and migration out of the IZ in slice cultures  regulate axon induction in vitro requires phospholipase Cγ, calcium, and calcium-calmodulin regulated kinase kinase . Dual inhibition of TrkB and TrkC or knockdown of phospholipase Cγ or CaMKKα inhibited axon outgrowth and radial migration in vivo, suggesting that signaling through phospholipase Cγ and CaMKKα is important. An alternative mechanism involves α2-chimerin  in progenitors inhibited axon outgrowth but neurons still migrated out of the IZ and into the CP, albeit in reduced numbers and with abnormal, highly branched leading processes  for Rap1, and increased GTP binding to Rap1   and Neuropilin (Nrp) family proteins are co-receptors for secreted Semaphorins, such as Sema3A. Importantly, Sema3A is secreted at the top of the CP and forms a gradient through the cortex. It is thus ideally suited to be a chemotactic factor for radial migration.In vitro, Sema3A attracts neurons when added to one side of a slice culture or to one side of a porous membrane  and gain of function  experiments to demonstrate necessity and sufficiency of specific proteins for radial migration. Some of the proteins, including LIS1 (Pafah1b1), Ndel1, dynamin, FilaminA and Doublecortin (DCX), were reviewed thoroughly previously, and will not be considered here  and Eph family proteins Figure .Efns are cell surface proteins that activate Eph family RTKs on other cells and generate “forward signals”. Reciprocally, an Eph can activate “reverse signaling” into an Efn-expressing cell. EfnA molecules are glycosylphosphatidylinositol-linked cell surface proteins that bind EphA family members. EfnB molecules are transmembrane proteins that bind EphB family members. Efns and Ephs regulate cell sorting in many developmental systems Fagotto, .EfnA and EphA genes are expressed in the developing cortex (Torii et al., Several Somewhat paradoxically, over-expression of an EfnA ligand, EphA7 or EphA4, caused over-expressing neurons to segregate from non-expressing neurons in the IZ (Torii et al., EfnB1 at E13 in post-mitotic neurons caused over-expressing neurons to segregate away from non-expressing neurons (Dimidschstein et al., P-Rex1. The abnormal clustering induced by over-expressing EfnB1 required the EfnB1 C terminus, P-Rex1 and Rac3, a little studied Rho-family member that is a primary substrate for P-Rex1 and is implicated in opposing Rac1 and reducing neurite number (Dimidschstein et al., In contrast with EfnAs stimulating lateral movement by forward signaling, endogenous EfnBs may inhibit lateral movement by reverse signaling. Over-expression of Time-lapse recordings and birthdating studies indicate that MP migration lasts for at least a day in the mouse, but the timing is very heterogeneous (Noctor et al., One major source of variability is that post-mitotic neurons and IP enter and leave the IZ at different rates (Tabata et al., One idea is that fates are not fixed at the last division of the RG progenitor but remain plastic (Fishell and Hanashima, in utero manipulation, and slice culture methods have led to the discovery of a large number of genes that regulate MP migration, axonogenesis, and direction of movement. Many of the genes have been subjected to epistasis analysis, implying causal relationships and sequences of contingent events. Some protein activities are regulated by external signals, while others may be “hard-wired” by cell intrinsic mechanisms or transcription changes. Despite dramatic progress, it is still unclear how these different processes fit together. We have limited understanding of how different pathways interact and exactly when and where in the cell signaling occurs. Analysis is extremely challenging given the asynchrony of the cell population, the small size of the cells, and the difficulties of imaging signaling events within living tissue. However, given the continuous development of new technology, we can be optimistic that these obstacles will be overcome.Amazing technical refinements in mouse genetics, imaging, The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Achieving sustainable agriculture and producing enough food for the increasing global population will require effective strategies to cope with harsh environments such as water and nutrient stress, high temperatures and compacted soils with high impedance that drastically reduce crop yield. Recent advances in the understanding of the molecular, cellular and epigenetic mechanisms that orchestrate plant responses to abiotic stress will serve as the platform to engineer improved crop plants with better designed root system architecture and optimized metabolism to enhance water and nutrients uptake and use efficiency and/or soil penetration. In this review we discuss such advances and how the generated knowledge could be used to integrate effective strategies to engineer crops by gene transfer or genome editing technologies. However, high input agriculture has been conducted with the excessive use of agrochemicals, including phosphorus (P)- and nitrogen (N)-fertilizers, herbicides, and insecticides. Moreover, the harsh conditions that crops face, such as drought, soil elemental toxicities, extreme temperatures, and high soil impedance, merit special attention because they drastically limit crop yields worldwide3. The predicted increase in the global population to over 9 billion by 20504 poses a critical challenge: how to develop effective stress-resistant/tolerant crops that are more competitive and can grow in marginal soils to ensure food production.Current agricultural systems use numerous crop varieties that have been improved through traditional breeding, which has produced a substantial increase in the yields of many crops, particularly cereals5. The transgenic crops that are currently on the market address the crop yield per unit area by controlling insect attacks and weed competition; however, new transgenic crops that overcome the limitations caused by harsh environments are also starting to be deregulated for commercial use5. In this review, we highlight some relevant transgenic approaches regarding nutrient use efficiency, abiotic stresses and soil physical degradation. These approaches have the potential to increase crop yields in marginal lands with poor soil fertility or low water availability and to expand cropping land into places in which the agro-climatic conditions are favourable but abiotic stress reduces yields and thereby discourages agricultural production.The development of efficient gene transfer systems, together with combined “omics” platforms , has facilitated the understanding of the physiology and biochemistry of plant adaptive responses to unfavorable environmental conditions and the identification of the key molecular players that control these responses. Several genes encoding transcription factors (TFs), transporters, and metabolic enzymes with a clear potential to improve crops have been identified. Agricultural schemes that use transgenic crops have proven to be effective and complementary alternatives because they provide multiple benefits for farmers ; such crops are cultivated today on more than 180 million hectares globally2PO4-/HPO4-2, Pi), whereas N is predominantly taken up as nitrate  or ammonia (NH4+)7. Moreover, the availability of Pi in the soil solution is drastically affected by the biogeochemical properties of the soil, making P-fertilization efficiency highly variable and more dependent on external inputs. To ensure high yields, farmers usually apply excessive amounts of both P- and N-fertilizers. This practice is unsustainable because crops use only 20–40% of the applied nutrients; the remainder contributes to environmental pollution, toxic algal blooms, and global warming8. Whereas N-fertilizers are synthesized from atmospheric N through a process that consumes at least 1% of global energy usage, P-fertilizers are produced from phosphate rock, a finite, non-renewable mineral resource. Consequently, both fertilizer and food prices will increase continuously. Therefore, searching for integrated strategies to increase P, N, and water use efficiency is an issue of food security and sustainability for all nations. The following paragraphs discuss the most relevant advances in engineering improved P and N uptake and use efficiency.Among all of the nutrients required by plants, P and N are the most limiting factors for agricultural production in most soils; thus, large amounts of fertilizers are commonly applied to ensure high yields. Although plants are able to use different organic compounds as sources of nutrients, P can be assimilated only in the form of orthophosphate  in rice, responsible for regulating the localization of low- and high-affinity Pi transporters to the plasma membrane21, results in enhanced low-Pi tolerance. Field data demonstrate that grain yield of PHF1-overexpressing plants in a low-Pi soil is higher than that of wild-type (WT) plants, suggesting that post-transcriptional regulation of Pi transporters could also be considered to improve crop performance in soils with low-Pi availability22.The uptake of N and Pi from the soil is critical and requires specialized transporter proteins25. However, except in the case of alanine aminotransferase (AlaAT)26, as described below, the overexpression of enzymes has not provided reproducible or robust results to indicate that it could be an effective strategy for improving the efficiency of N use.The generation of transgenic plants to improve the N use efficiency has also been attempted in a variety of crop plants by manipulating the flux-limiting enzymes involved in N assimilationArabidopsis, and it is evolutionarily conserved from algae to vascular plants27. Overexpressing Phosphate Starvation Response 1 (PHR1) and other TFs, such as Phosphate Starvation-Induced Transcription Factor 1 (PTF1) and OsMYB2P-1, in a variety of crops, such as wheat28, rice29, and maize30, appear to confer low-Pi tolerance and improved grain yield in greenhouse or field trials. Field-testing in different geographical locations and different soil types is required to confirm that the overexpression of these TFs is a robust strategy for improving plant performance under Pi-limiting conditions without affecting performance under optimal Pi availability. Recently, it was reported that inArabidopsis and rice, SPX1 and SPX2 repress the activity of PHR1 as a transcriptional activator in a Pi-dependent manner32. The data published in these reports strongly suggest that the PHR1-SPX1/SPX2 complex is one of the main sensors that regulate the plant response to low-Pi availability. Although no structure of the PHR1-Pi-SPX complex is available, regulation of the interaction between these proteins could become an important target for engineering plants with modulated responses to low-Pi availability. It could be possible to alter PHR1 or SPX1/SPX2 in such a way that the low-Pi response could be modulated to activate processes that enhance Pi uptake and assimilation while preventing the drastic reduction in shoot growth that is generally observed in Pi-starved plants  TF was recently identified as a key element in the systemic signaling pathway that directs N foraging inArabidopsis roots, thus opening up the possibility of controlling root plasticity to improve soil exploration capacity in crops34.In the case of N, overexpression of the TaNFYA-B1 TF in wheat improved the yield under different regimes of P and N inputs under field conditions35. These enhancer elements could be used to design synthetic promoters that could direct high levels of expression while maintaining cell specificity and responsiveness to Pi- or N-deprivation. An additional phase for the improvement of nutrient uptake and use efficiency will be the understanding of the regulatory networks that orchestrate plant responses to nutrient deficiency. The integration of this knowledge will serve to design strategies to direct the enhanced expression of two or more TFs simultaneously, leading to more robust improvements in the key traits to achieve a more sustainable agriculture  and marker-assisted backcrossing of genes that regulate root traits that improve nutrient uptake and use efficiencies into modern varieties is of the highest importance26, the use of PHOSPHORUS-STARVATION TOLERANCE 1 (PSTOL1) to enhance P assimilation41, and the development of a novel fertilization system based on the production of transgenic plants that are able to use phosphite (Phi) instead of Pi as a P source42  plays an important role in carbon fixation and N metabolism because it catalyzes the reversible reaction between pyruvate and glutamate to produce alanine and oxoglutarate43. The potential effectiveness of this approach relies on the facts that amino acids act as signals controlling N uptake and that alanine is the only amino acid whose biosynthesis is not inhibited by N deficiency43. The development of the AlaAT technology started with the expression of barley AlaAT in canola using thebtg26 root-specific promoter, which resulted in the production of increased biomass under low-N conditions26. Field evaluations showed thatbtg26:AlaAT canola plants exhibited a 42% increase in seed yield under suboptimal N fertilization (56 kg ha-1). This yield increase correlated with lower levels of glutamine and glutamate in the shoot, increased N influx and higher levels of alanine in roots, and up-regulation of high-affinity N-transporters. The overexpression of barley AlaAT in rice, which was also driven using a root-specific promoter (OsANT1), also resulted in increased biomass and grain yield44. Recently, similar results have been reported for sugarcane45.btg26 promoter is expressed mainly in the cortex and lateral roots of transgenic plants26, which are fundamental for the uptake and loading of nutrients into the vascular system. It will be interesting to determine how robust the AlaAT-overexpressing phenotype is under different stress conditions and soil types.Interestingly, thePhosphorus-starvation tolerance 1. Because of the low mobility of Pi in the soil7, a well-developed and highly branched root system is a determinant for soil exploration and Pi uptake in soils with a low availability of this nutrient46. However, modern breeding programs have focused on developing high-yield crops by primarily selecting the above-ground phenotype and applying full fertilization during breeding processes, which probably selects against root traits that are important for nutrient uptake efficiency. The QTLPup1 (Phosphorus Uptake1), which is responsible for low-Pi tolerance, was identified in a cross between a low-Pi-tolerant rice landrace with a low-Pi-intolerant modern rice variety41.Pup1 was found to contribute to enhanced Pi uptake and grain yield by 170% and 250%, respectively, in low-Pi soils in the modern variety47. Recently, it was found that thePSTOL1 gene, which encodes a protein kinase, is responsible for the effect of thePup1 QTL on Pi uptake and assimilation41. The overexpression ofPSTOL1 under a constitutive promoter (CaMV35S) in two types of modern rice varieties (one indica and one japonica) that naturally lack the gene resulted in an increase of over 60% in grain yield in low-Pi soils41.41. Because peptide transporters are included in the set of N-transporters (PTR/NRT1) in plants48, it would be interesting to determine whetherPSTOL1-overexpression could also improve N uptake efficiency. Optimizing the cell-specific and regulated expression ofPSTOL1 will probably have an even higher impact on grain yield.Interestingly, the low-P tolerance phenotype conferred by PSTOL1 correlated with a more robust root system, as transgenic plants produced almost five times more root biomass than did the non-transgenic plants. A global expression analysis of the PSTOL1-overexpressing plants revealed a set of up-regulated genes that are related to root growth and stress responses, including a putative peptide transporterThe phosphite oxidoreductase/phosphite system. The high reactivity of Pi with soil components and the constant competition of microorganisms and weeds with cultivated plants make agriculture highly dependent on P-fertilizers and herbicides. Recently, a phosphite oxidoreductase (PTXD) fromPseudomonas stutzeri was used to propose a re-design of the currently used agricultural systems42. PTXD oxidizes Phi using NAD+ as a cofactor and yields Pi and NADH as products49. The expression of PTXD inArabidopsis and tobacco produced transgenic plants that are capable of using Phi as a sole P source.50; in addition, plants and most microorganisms are unable to metabolize Phi as a P source53. Therefore, the system makes the PTXD-transgenic plants more competitive over other plants, including weeds, in low-Pi soils that are fertilized with Phi. PTXD-transgenic lines required 30–50% less P to achieve optimal productivity when they were fertilized with Phi instead of Pi42 and reduced the requirement of herbicides because of the poor growth of weeds in soils fertilized with Phi.The importance of this approach relies on the fact that Phi has distinct chemical and biochemical properties compared with Pi, including higher solubility and lower reactivity with soil componentsThis technology is still in its infancy, and several questions need to be addressed before its real potential is uncovered. The promising results obtained under greenhouse and field conditions suggest that a potential reduction in production costs and energy consumption could be achieved by replacing the independent application of fertilizer and herbicides with a single treatment and by reducing the cost of additional herbicides. It is important to consider that the PTXD/Phi system has the highest potential for use in acidic and alkaline soils that have very low-Pi availability and that, in spite of having appropriate climatic conditions and water availability to sustain high crop productivity, it has been used only as grassland for cattle. These areas are predominant in Brazil, China, Australia, India, and Russia, comprising over 350 million hectares that could be converted into highly productive cropping areas if the PTXD/Phi technology was incorporated into genotypes adapted to acidic and alkaline soils.54.This technology provides sustainable management of P, and thus has the potential to prolong the lifetime of phosphate rock reserves and to reduce the environmental impact of eutrophication of lakes, seas, and oceans. Additionally, a paper published recently suggested that on the moon or other planets that lack oxygen in the atmosphere, on which P accumulates primarily as schreibersite mineral and that are rich in Phi, the PTXD/Phi technology could be an interesting alternative for establishing agriculture3+ ions are dissolved from soil minerals and are highly toxic to plants, impairing root growth and function. Two main classes of Al resistance mechanisms have been reported: Al exclusion mechanisms, which prevent Al from entering the root apex, and Al tolerance mechanisms, in which Al enters the plant but is sequestered into the vacuole and detoxified. Since the root apex is the main site of Al toxicity, the most well-characterized exclusion mechanism involves the regulated release of organic acids (OAs) by the root tip, which chelate Al3+ ions forming non-toxic compounds that do not enter the root tip cells. Members of the Al-activated Malate Transporter (ALMT) family of anion channel transporters and the Multidrug and Toxic compound Extrusion (MATE) family of OA/H+ antiport transporters are responsible for plasma membrane malate and citrate efflux, respectively, from root cells into the rhizosphere in response to the presence of toxic concentrations of Al3+ ions  and boron (B) toxicity. Al toxicity is a major constraint for plant yield on acidic soils, which comprise between 40 and 50% of the world’s potentially arable lands. At pH values below 5, AlBot1 (encoding a B efflux transporter), which is highly expressed in the tolerant landraces58, whereas B tolerance in wheat is associated with a B transporter-like gene (Bot-B5b) that has high root expression levels in tolerant genotypes as compared to susceptible lines59. The finding that high expression of B exporters reduces B concentration in the plant, or that a decreased expression of the transporters that facilitate B uptake could lead to tolerance to toxic B levels, opens up the possibility of using transgenic approaches or genome editing technologies to improve the yield of different crops in soils containing toxic levels of B.B is an essential micronutrient required for several physiological and developmental processes in plants, including meristem development, but that can also be present in toxic levels in the soil. Typical B toxicity symptoms include necrosis of marginal leaves and the inhibition of root growth (for a review see60.61. Heat shock proteins (HSPs) and late embryogenesis abundant (LEA) proteins also play crucial roles during seed desiccation and water stress by preventing protein denaturation and aggregation62. Additionally, several genes that encode TFs have been identified as key elements that possess the potential to improve crop performance under different abiotic stresses  andCspB genes fromEscherichia coli andBacillus subtilis, respectively. These genes encode RNA-binding proteins with chaperone activity that confer drought tolerance in maize and rice under field conditions67. In fact,CspB-expressing maize is the first genetically modified (GM) crop with enhanced water use efficiency that has been deregulated for commercial use in the USA5.HSPs and LEA proteins from plants have been clearly shown to be involved in abiotic stress responses; however, as shown instrated . Interes69. To develop crops with higher yields under drought, it will most likely be necessary to engineer metabolic pathways through the simultaneous manipulation of multiple critical genes. In addition, it would be interesting to explore the mechanisms that regulate desiccation tolerance in seeds to obtain new insights into the adaptive stress response pathways and to identify new candidate genes for crop improvement.The multiple pathways involved in plant adaptations to osmotic and water stress and the complexity of their interactions can explain, to some extent, the limited success under field conditions of manipulating individual genes encoding chaperones or enzymes involved in the synthesis of osmoprotectants70, NAM-ATAF and CUC (NAC)71, and Nuclear Factor Y (NF-Y)72 families have been used to develop transgenic plants and study their performance under stress conditions. The expression of some of these TFs under drought-inducible or root-specific promoters has resulted in improved tolerance to drought, salinity, and temperature stress and a higher yield under water-limited conditions in rice76, wheat77, canola78, and maize72.Manipulating proteins that regulate gene expression or the signal transduction of multiple metabolic pathways involved in abiotic stresses has proven to be useful for improving the stress tolerance of crops . TFs tha79 to identify and characterize the Deeper Rooting 1 (DRO1) QTL that controls the root growth angle80. Higher expression of DRO1 causes a more vertical root growth. Breeding DRO1 into a shallow-rooting rice line enables these plants to avoid drought by increasing the depth of their roots, resulting in a higher grain yield80. The DRO1 gene is the first drought tolerance QTL that was cloned, and its beneficial effects on plant growth further confirmed that the root system architecture plays a crucial role in abiotic stress tolerance. Interestingly,DRO1 has no homology to known proteins, which suggests that cloning genes associated with QTLs could provide completely novel genes for plant breeding. This example shows that a considerable improvement in drought tolerance can be achieved by altering root growth patterns and opens up the possibility of introducing DRO1 in shallow-rooting crops other than rice through the use of genetic engineering  breedingneering .82. This suggests that the precise regulation of the activity of individual or multiple receptors will be required to achieve enhanced drought tolerance without a yield penalty. A novel alternative to actively control tolerance to abiotic stress is the use of chemicals that can activate or repress the receptors that sense the stress or the signaling pathways activated by hormones that mediate the corresponding stress responses. Recently it was shown that it is feasible for the case of drought tolerance. Drought tolerance inArabidopsis was achieved using an engineered PYR1 ABA-receptor that can be activated by an existing non-herbicidal agrochemical that is not a natural inducer of ABA responses. This example opens up a new avenue of crop improvement to regulate abiotic or biotic stress responses at the beginning of, or prior to, the presence of the stress in a timely and quantitative manner by the application of a non-toxic compound, reducing potential yield reductions83.The phytohormone abscisic acid (ABA) regulates numerous processes in plants including seed dormancy and the plant responses to low water availability. ABA is perceived by soluble PYR/PYL/RCAR (pyrabactin resistance1/PYR1-like/regulatory component of ABA receptor) receptors that belong to the START superfamily of ligand-binding proteins , and transposable element activity, play essential roles in modulating gene activity in response to environmental stimulid crops . However108. Furthermore, the possibility of activating or repressing the expression of specific genes by introducing site-specific epigenetic changes, such as DNA methylation or histone modifications using a modified version of the CRISPR/Cas9 system99, will drastically modify how agriculture is developed by creating an integral, effective, and sustainable global agriculture. However, translating these approaches from the laboratory or the greenhouse to the field remains challenging. In our opinion, more interdisciplinary research and the active involvement of breeders and agronomists in project planning is necessary to better define project goals and align the interests of researchers with that of crop producers.We believe that the most exciting transgenic approaches for producing plant varieties and hybrids that are much less dependent on the application of agrochemicals, including fertilizers and pesticides, have yet to be discovered. The engineering of crops for harsh environments is evolving and will rapidly incorporate new breeding technologies, including genome editing, which has already produced its first commercial product (herbicide-resistant canola). The development of effective approaches for specifically and visibly monitoring certain environmental stresses, such as P deficiency, and timely indicating the degree of the stress is also emerging and providing additional tools for improving crops I have read this submission. I believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. I have read this submission. I believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.


Whilst there is compelling evidence of an almost 2-fold increased risk of still births, and suggestive evidence of increased mortality among offspring of mothers with psychotic disorders, only three studies have addressed the role of antenatal depression (AND) on survival of the baby. We examined these associations in a large cohort of pregnant women in Ghana.A Cohort study nested within 4-weekly surveillance of all women of reproductive age to identify pregnancies and collect data on births and deaths in the Kintampo Health Research Centre study area of Ghana. Women were screened for AND using the Patient Health Questionnaire (PHQ-9) to ascertain DSM-IV major or minor depression. Outcomes were adverse birth outcomes, maternal/infant morbidity, and uptake of key newborn care practices, examined using logistic regression; effect sizes reported as relative risks with 95% confidence intervals.20679 (89.6%) pregnant women completed the PHQ-9. The prevalence of AND was 9.9% (n = 2032) . AND was associated with: prolonged labour ; peripartum complications ;postpartum complications ; non-vaginal delivery ; newborn illness ; and bed net use during pregnancy , but not neonatal deaths, still births, low birth weight, immediate breast feeding initiation, or exclusive breastfeeding. AND was marginally associated with preterm births .This paper has contributed important evidence on the role of antenatal depression as a potential contributor to maternal and infant morbidity. Non-pharmacological treatments anchored on primary care delivery structures are recommended as an immediate step. We further recommend that trials are designed to assess if treating antenatal depression in conjunction with improving the quality of obstetric care results in improved maternal and newborn outcomes. Global efforts to reduce the burden of neonatal deaths appear to be yielding positive results with a 1.7 per annum decline rate since 1990 Whilst there is compelling evidence of an almost 2-fold increased risk of still births , and suggestive evidence of increased mortality among offspring of mothers with psychotic disorders There is, however, strong evidence linking antenatal depression and other adverse birth outcomes such as preterm births  and low birth weight  as reported in a recent meta-analysis involving 26 studies from high income settings and three from low income settings This paper presents findings from a large cohort study conducted in Ghana to address the relative lack of evidence from low and middle income settings, particularly SSA concerning the burden, determinants, and adverse consequences of perinatal depression. The findings presented in this paper include the association of antenatal depression with both adverse birth and maternal morbidity outcomes including birth complications, prolonged labour, and assisted delivery. Finally we present associations with uptake of key newborn care practices.Depression nested within the ObaapaVitA Newhints DON is a cohort study of antenatal and postnatal DON was carried out from late January 2008 to early August 2009 and comprised depression assessments in the 4-weekly surveillance visits following identification of pregnancy and in the visits following reporting of a delivery. The analysis in this paper focuses on the consequences of antenatal depression for the mother and baby.All women of reproductive age were visited at home every 4-weeks by a locally-resident field worker, in order to collect self-reported data on pregnancies, deliveries, including morbidity. When a pregnancy was first reported, information was collected on: socio-demographic and socio-economic indicators, including obstetric history. A DON pregnancy depression assessment was then conducted at the following 4-weekly visit using the Patient Health Questionnaire (PHQ-9). At the first visit after the delivery was reported , information was collected on the pregnancy, delivery, any complications, the baby (or babies), and new born care practices.The assessments of antenatal depression were made by administering the Twi (widely spoken language in Ghana and the study area) version of the 9-item Patient Health Questionnaire (PHQ-9) The PHQ-9 is a short structured questionnaire that enquires about the nine symptom based criteria for a diagnosis of Diagnostic and Statistical Manual version four (DSM-IV) We examined three main types of outcomes and these were all reported by the mother at the next 4 weekly visit to the mother after birth. The first is adverse birth outcomes, which covered: neonatal deaths and still births, preterm deliveries (based on the mother’s report on whether the delivery was less than 4 weeks (<37 weeks) before the due date), and any illness that the mother had thought was serious or severe, and low birth weight (LBW) (<2.5 kg). The preterm births (PTB) reported in this study are a result of spontaneous preterm labour. Data collectors were trained to ask this question carefully in order to rule out other possible explanations for the PTB. Birth weight was extracted by fieldworkers from the birth cards given to mothers who delivered in facilities; it was not therefore available for babies delivered at home.The second type of outcome is maternal morbidity outcomes pertaining to the birth. Women were asked about serious problems they may have experienced during labour/soon after birth or since birth. These were: assisted deliveries , prolonged labour (23+hrs), peripartum complications , and postpartum complications , smelly vaginal discharge, leaking urine/faeces, mastitis, and any other problem not mentioned by the field worker). Data collectors were trained to enquire after these experiences with relevant examples. For example, mastitis is explained as a breast infection that is swollen, painful, or has a discharge, also known as ‘pompo’ in the local language. The list of questions was based on standard maternal morbidity questions which were adapted and piloted in the year 2000 when the surveillance system was established.The third type is uptake of key newborn care practices: attending at least 4 antenatal care sessions, initiating breast feeding within an hour of birth, exclusively breast feeding within the first month based on the mother’s account of breast feeding in the last 24 hours.A priori potential confounders were: maternal characteristics ; pregnancy and obstetric variables (parity). In addition, an overall socio-economic ‘score’ for each woman was generated using factor analysis techniques after the methods described by Vyas and Kumaranayake (2006) Over 20,000 pregnant women were screened for depression. With the prevailing rates of neonatal mortality and still births within the study population, coupled with the sample sizes available and antenatal depression prevalence, we estimated to be able to detect an effect size of 1.23 with 95% confidence intervals and 80% power.Analyses were based on women who had a DON depression screening during pregnancy, and their babies, and were restricted to singletons. Multiple pregnancies were excluded from all analyses given the high risk associated with infant mortality particularly in Africa Ethical approval for the study was granted by the ethics committees of the Kintampo Health Research Centre (KHRC) and the London School of Hygiene and Tropical Medicine. Written informed consent was obtained from all participants using an informed consent form that was approved by the ethics committees. The informed consent procedure was either conducted in English or the local language (Twi) for those women who were not literate in English; in such instances a literate witness was involved.rd December 2007 and 25th June 2009, 26,980 pregnant women were identified, of whom 23,011 were eligible for depression assessment. Of these 20,679 (89.8%) pregnant women with a singleton birth (live/still) completed the depression screen. This is the denominator used for analyses examining maternal morbidity outcomes and pregnancy behaviours, and still births. Forty-three (0.2%) declined to participate, 1463 (6.4%) were temporarily absent at the surveillance visit, and 370 (1.6%) did not have a depression form completed although they were visited. Background characteristics of those not met and screened were comparable to those in the analysis  . Detailed profile of study participants is given in a companion paper This is the largest cohort study that has yet been conducted in low income settings of the effects of antenatal depression on adverse outcomes for the mother and baby. After adjustment for confounders, antenatal depression was not found to be associated with neonatal deaths, still births, low birth weight, delayed initiation of breastfeeding, or non-exclusive breastfeeding in the neonatal period, delivering at a health facility, or optimal antenatal care attendance. However, antenatal depression was associated with a 25% increase in 24+ hours prolonged labour, 11% severe peripartum and 27% postpartum complications, 50% severe newborn illness, and 7% less bed net non-use during pregnancy. It was also marginally associated with a 32% increase risk of preterm deliveries.Our report is strengthened by several factors. We employed an unprecedentedly large cohort of 20,679 pregnant women, applied clinimetric criterion for depression using a locally validated tool, ascertained outcomes blind to exposures because the data collectors were not aware of the study hypothesis, and recorded a high antenatal depression screening response rate of 92%. Further to this, we employed a more robust ascertainment of neonatal deaths based on the frequent 4-weekly surveillance visits to mothers. Our measure of low birth weight is also robust as these were recorded at the health facility.Possible weaknesses are our self-reported morbidity outcomes may have been influenced by the mother’s depression status after birth; however this may not have had the assumed biased effect as 87% of women depressed antenatally will usually not be depressed at their postpartum assessment in this setting In addition our measure of antenatal depression was not originally validated in the antenatal period and given that one study suggests depression in pregnancy and postnatal period show significantly different symptom profiles Our finding of an independent association between poor antenatal mental health and prolonged labour is consistent with findings from both LMIC The evidence on the association between antenatal depression and poor birth outcomes including the survival of the baby in low and middle income settings is scanty. We are aware of only one study in SSA (Ethiopia) that examined the association between antenatal depression and neonatal mortality, and this showed no evidence of any increased risk Our negative finding on LBW, is in contrast to the increased risks found in other cohort studies in developing countries in Ethiopia Our study did not also replicate the positive association between antenatal common mental disorder and delayed initiation of breastfeeding in Ethiopia  Though perinatal depression has been shown to affect the health-related quality of life of the mother Although antenatal depression in our setting and SSA in general may be self-limiting Antenatal depression is associated with:Low birth weight/stunted growth particularly in south AsiaPreterm birthsProlonged labourPostnatal depressionPoor maternal self-care and nutritionAntenatal depression is associated with:Severe newborn illnessBed net non-use during pregnancySevere peripartum and postpartum complicationsInstrumental/caesarean sectionS1 TableEffect of antenatal depression on risk of specific peripartum complications.(DOCX)Click here for additional data file.S2 TableEffect of antenatal depression on risk of specific postpartum complications.(DOCX)Click here for additional data file.


Caloric restriction is the most effective and reproducible dietary intervention known to regulate aging and increase the healthy lifespan in various model organisms, ranging from the unicellular yeast to worms, flies, rodents, and primates. However, caloric restriction, which in most cases entails a 20–40% reduction of food consumption relative to normal intake, is a severe intervention that results in both beneficial and detrimental effects. Specific types of chronic, intermittent, or periodic dietary restrictions without chronic caloric restriction have instead the potential to provide a significant healthspan increase while minimizing adverse effects. Improved periodic or targeted dietary restriction regimens that uncouple the challenge of food deprivation from the beneficial effects will allow a safe intervention feasible for a major portion of the population. Here we focus on healthspan interventions that are not chronic or do not require calorie restriction. Here we discuss more recent discoveries that have advanced our knowledge of the effects of less invasive and restrictive dietary interventions in aging and healthspan.Aging is the leading risk factor for many among the major diseases and a key factor in the overall decline of physical and mental performance. Interventions that slow down the aging process can delay or prevent multiple chronic diseases and improve productivity and quality of life in older ages. For clarity, here, we use caloric restriction (CR) to refer to a dietary intervention with an overall 20–40% reduction of total caloric intake, and dietary restriction to represent a broader scope of dietary interventions that encompass those with specific macronutrient and feeding pattern restrictions. CR is the most studied and reproducible non-genetic intervention known to extend healthspan and/or lifespan in organisms, ranging from unicellular yeast to monkeys. It started with a simple experiment where a reduction in dietary intake  extended the lifespan of rats1, providing an experimental model to begin to demonstrate that aging can be slowed down. Nearly half a century later, Walford and Weindruch reported that “adult-initiated” caloric restriction started at 12 months of age not only increased lifespan but also reduced the incidence of spontaneous cancer by more than 50% in rats3. Several decades later, the effect of caloric restriction on healthspan and lifespan has been confirmed in model organisms ranging from unicellular yeast to worms, flies, rodents, and primates4, suggesting a highly conserved effect which may involve common genes. Although the molecular mechanisms that mediate the effect of caloric restriction are still being investigated and debated, there is more widespread acceptance of the hypothesis that caloric restriction and lifespan extension involves the down-regulation of insulin and insulin-like signalling (IIS)5, as well as of the amino signalling target of rapamycin (TOR)-S6 kinase pathway7, and the glucose signalling Ras-protein kinase A (PKA) pathway9.In their seminal report in 1935, Crowell and McCay demonstrated that simply reducing caloric intake without causing malnutrition nearly doubled the lifespan of rats12. In yeast, down-regulation of (a) the amino acid-sensing TOR and the ribosomal protein S6 kinase (S6K) ortholog Sch9 pathway6, and (b) the Ras-AC-PKA pathway13 are key changes mediating part of the effects of caloric restriction on chronological lifespan, the measurement of cellular survival under non-dividing conditions. In contrast, elevated activity of sirtuin (SIR2) has been described as a key change in the extension of replicative lifespan, measured by counting the number of buds generated by an individual mother cell15. In worms, the lifespan extension caused by the inactivation of IIS, or by different forms of caloric restriction, requires Forkhead FoxO transcription factor daf-1616. In flies, the IIS pathway is involved in the effects of caloric restriction17 and, although dFoxo is not required for its longevity effect, its activity can affect the response to caloric restriction18. In rodents, growth hormone (GH) and IGF-1 levels are reduced following caloric restriction19, but the link between dietary restriction, GH and aging is still being investigated, with focus on the genes and pathways regulating longevity in the simple organisms described above. The long-lived GH receptor knock-out (GHRKO) mice, which are resistant to GH, do not exhibit further lifespan extension or health benefits by caloric restriction21, but the long-lived GH-deficient Ames mice do22, suggesting a complex involvement of the GH/IGF-1 axis and periphery pathways in the response to caloric restriction.Because an in depth discussion of the anti- and pro-aging pathways conserved in model organisms has been covered elsewhere, here we will only mention the most relevant ones23, whereas WNPRC reported significant improvement in both lifespan and healthspan24. The discrepancies are largely attributed to the different dietary composition and heterogenic genetic background23, which have been shown to be a significant factor in rodents26. This underscores the importance of diet composition and genetic background and their compatibility when applying caloric restriction to humans. Nonetheless, several studies provide evidence supporting beneficial health effects of caloric restriction for humans. A notable NIH-sponsored controlled randomized study on non-obese individuals, CALERIE (Comprehensive Assessment of the Long-term Effects of Reducing Intake of Energy), recently reported that a two year 25% caloric restriction is feasible for humans and provides health benefits, such as reduced inflammatory markers and cardiometabolic risk factors29. However, caloric restriction was associated with reduced bone mineral density and exercise was recommended to offset such adverse effects30. Notably, CALERIE was conducted in three independent centers and involved 218 overweight participants, suggesting that caloric restriction can be beneficial even in a very genetically heterogeneous group31. However, considering the results in monkeys, much longer and larger studies will be needed to know what the effects of CR on human healthspan.Much has been learned about caloric restriction and aging from model organisms, but the ultimate question that lingers is the relevance of these models to human lifespan and healthspan. The rhesus monkeys are the closest model organism to humans in which caloric restriction has been experimentally tested in a controlled environment. Two notable studies performed by independent programs, the National Institute on Aging (NIA) Intramural Research Program and the Wisconsin National Primate Research Center (WNPRC), subjected male and female rhesus monkeys to 30% caloric restriction from levels of baseline caloric intake. The NIA reported no improvement in lifespan but observed a positive trend for the delay of age-related diseases 32. An alternative hypothesis suggests that caloric restriction acts as a mild stressor that promotes hormesis, which refers to the beneficial effects resulting from the cellular responses to mild, repeated stress33. Stress resistance should also be considered an important criterion for the successful development of caloric restriction-mimetic dietary and pharmacological interventions.Among the cellular alterations most closely associated with both caloric restriction and longevity mutations is the resistance to multiple stressors, which in most cases includes resistance to oxidative stress. The ability of caloric restriction to prevent the damage caused by exogenous toxins is likely to be associated with the protection, repair and replacement effects that prevent the age-dependent dysfunction caused by endogenous processes and toxic molecules19. Both intermittent and periodic fasting can increase lifespan, even when there is little or no overall decrease in calorie intake50.The definition of dietary restriction has been expanded from an alternative description of caloric restriction to also encompass a broader scope of interventions, including short-term starvation, periodic fasting, fasting-mimetic diets, intermittent fasting, normo-caloric diets with planned deficiencies , and time-restricted feeding. Most of these relatively novel interventions are reported to have beneficial effects on overall health and in some cases longevity. Fasting is an extreme dietary intervention describing either a complete lack of food intake or a 60% or higher food restriction. Intermittent fasting refers to practicing this intervention every other day whereas periodic fasting refers to severe restriction for two or more days periodically . Caloric restriction and fasting share similar but often distinct effects on a number of biomarkers  suggesting that partially overlapping mechanisms are involved36. Simply reducing protein intake can deliver an equally potent impact on lifespan as dietary restriction in multiple model organisms35. A recent analysis of the National Health and Nutrition Examination Survey (NHANES) showed that low protein intake was associated with reduced overall mortality for those under 65 years of age37. Also, a high-carbohydrate, low-protein diet resulted in longer lifespan and improved cardiometabolic health, despite increased food intake and body fat39. Furthermore, the restriction of a single essential amino acid in a normal diet increased lifespan and stress resistance44. In flies, adding back essential amino acids to the caloric restriction diet decreased lifespan to that of the normally fed group36. Laboratory rodents fed a methionine-restricted diet displayed an extended lifespan with decreased age-dependent diseases and increased resistance to oxidative stress, in part due to increased antioxidant capacity48. A tryptophan-restricted diet also provided longevity and reduced age-dependent deterioration49 but has mainly been explored for neurological benefits, due to its role in serotonin synthesis. A fasting-mimicking diet, consisting of very low calorie and protein that leads to similar physiological response to fasting, including reduced levels of glucose and IGF-1 and increased levels of ketone bodies IGFBP-1, enhanced healthspan and rejuvenated the hematopoietic system while promoting adult neurogenesis50. Further studies on carbohydrate and fat restriction are needed to determine their role in dietary restriction.In addition to periodic or intermittent fasting-based strategies as alternatives to caloric restriction, the restriction of specific macronutrients (or macronutrient restriction) without the restriction of calories is among the most promising interventions that have emerged to promote healthy aging in humans. Among the different types of macronutrient restriction, reduced intake of proteins and amino acids is the most effective pro-longevity intervention53. When mice were given access to food for only 8–9 hours during the active phase of the day, metabolic diseases induced by a high-fat, high-fructose, and high-sucrose diet, were reduced without lowering caloric intake51. The benefits of time-restricted feeding against such obesogenic diets were proportional to the duration of the fasting each day52.Ad lib feeding during the weekend did not interfere with the protective effects of time-restricted feeding52. Notably, the restricted feeding pattern reversed the progression of pre-existing obesity and type II diabetes, suggesting it has the potential to be a clinically relevant and feasible dietary intervention, useful to prevent and treat obesity and metabolic disorders52. Considering that key metabolic factors, such as 5' AMP-activated protein kinase (AMPK), sirtuins, and protein kinase B (AKT), are regulated by an interplay of circadian rhythm and feeding time55, dietary schedules should be more carefully studied in the context of dietary restriction.Feeding schedule has also been shown to have a significant impact on health and survival. In flies, time-restricted feeding (limited to 12 daytime hours every day) had profound effects on neural, peripheral, and cardiovascular physiology and improved sleep, body weight maintenance, and delayed signs of cardiac aging, under unchanged caloric intake and activityDietary interventions that extend healthspan and lifespan have evolved and have become much better characterized since the original caloric restriction experiments performed by McCay in 1935. We now understand that its effects on aging are not simply the result of the reduced amount of calories consumed, but are also determined by diet composition, and can be achieved by periodic interventions which do not require an overall reduction in calorie intake and which can be achieved without a complete lack of food intake during the periodic fasting cycles. Further studies are important to identify even less invasive and more effective dietary interventions that will cause coordinated and beneficial effects on healthspan.


The Cytokine Storm Syndrome (CSS) is characterized by an overwhelming activation of immune cells observed in life-threatening disorders such as familial hemophagocytic lymphohistiocytosis (fHLH) and secondary (s) HLH/macrophage activation syndrome (MAS) as well as during serious infection. However, it is not known if the CSS can be attributed to a single cytokine. Increased blood levels of interferon gamma (IFNg) in HLH and sHLH/MAS patients potentially indicate a central role for this cytokine in the CSS. Using a mouse model that mimics an infection-driven CSS , our study showed that total IFNg levels originating within organs are 500 to 2,000-fold higher than those measured in peripheral blood as CSS develops. Ablation of IFNg activity in tissues led to the amelioration of the plethora of associated CSS clinical and laboratory parameters. Furthermore, the IFNg signature gene products, CXCL9 and CXCL10, correlated with disease severity in the mouse model of CSS and patients with sHLH. Thus, anti-IFNg targeted therapy should control diseases associated with the cytokine storm and we propose the use of CXCL9 or CXCL10 as a means to monitor total IFNg activity in patients.


The etiology of amyotrophic lateral sclerosis (ALS), a fatal motor neuron disorder characterized by progressive muscle weakness and spasticity, remains largely unknown. Approximately 5–10% of cases are familial, and of those, 15–20% are associated with mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). Mutations of the SOD1 gene interrupt cellular homeostasis and contribute to cellular toxicity evoked by the presence of altered SOD1, along with other toxic species, such as advanced glycation end products (AGEs). AGEs trigger activation of their chief cell surface receptor, RAGE (receptor for advanced glycation end products), and induce RAGE-dependent cellular stress and inflammation in neurons, thereby affecting their function and leading to apoptosis. Here, we show for the first time that the expression of RAGE is higher in the SOD1 transgenic mouse model of ALS vs. wild-type mouse spinal cord. We tested whether pharmacological blockade of RAGE may delay the onset and progression of disease in this mouse model. Our findings reveal that treatment of SOD1 transgenic mice with soluble RAGE (sRAGE), a natural competitor of RAGE that sequesters RAGE ligands and blocks their interaction with cell surface RAGE, significantly delays the progression of ALS and prolongs life span compared to vehicle treatment. We demonstrate that in sRAGE-treated SOD1 transgenic animals at the final stage of the disease, a significantly higher number of neurons and lower number of astrocytes is detectable in the spinal cord. We conclude that RAGE antagonism may provide a novel therapeutic strategy for ALS intervention. Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig's disease, is a progressive and terminal neurological disorder, which results in severe disability and death within 5 years after diagnosis , and interacts with a number of other molecules such as S100/calgranulin family members, High Mobility Group Box-1 (HMGB1), and molecules that are prone to aggregation and post translational modifications  is a soluble form of RAGE lacking the transmembrane region, composed of the extracellular domains of the receptor. sRAGE acts as a RAGE decoy, thereby inhibiting RAGE function and lowering levels of RAGE ligands, effectively preventing harmful RAGE ligand effects on cells Schmidt, . sRAGE cIn the present study, we tested for the first time whether sRAGE treatment offers protection against premature cell death in motor neurons of SOD1 transgenic mouse, thereby delaying the onset and progression of ALS and improving overall health status of these mice. Our findings demonstrate the upregulation of RAGE in the spinal cord of SOD1 mice, and show that sRAGE-treated SOD1 mice display improved lifespan, improved functional motor performance scores, and less neuronal cell death compared to vehicle-treated age-matched SOD1 transgenic mice controls. The results of this first study testing RAGE as a therapeutic target in ALS support further examination of this receptor in this incurable disorder.*G93A) 1Gur/J mice of both genders  were used in the study. According to the Jackson Laboratories site (http://jaxmice.jax.org/strain/002726.html), transgenic mice have an abbreviated life span: 50% survive at 128.9 ± 9.1 days .Quantification of lumbar ventral horn motor neurons and glial cells was performed on 4% paraformaldehyde-fixed end-stage spinal cords from MSA- and sRAGE-injected SOD1 mice. For neuronal counts, 10 μm thick sections of lumbar spinal cord were cut serially on the cryostat and stained with Cresyl Violet following standard histological protocol. For astrocytosis counts, 10 μm thick spinal cord sections were stained with primary rabbit anti-GFAP antibody  followed by staining with biotinylated secondary antibodies according to the standard peroxidase anti-peroxidase (PAP) protocol provided with the kit . For each condition, 25 sections were studied, five sections per one tissue sample, 50 μm between sections within each tissue sample. Representative regions of interest, 200 × 200 μm, outlining ventral horn regions were analyzed and the same region was analyzed in each sample. All sections were examined with the Leica microscope  and quantified using Image J analysis tool  and cDNA was synthesized with iScript cDNA Synthesis Kit . Quantitative Real Time PCR for the gene encoding mouse RAGE, Ager, was performed using the TaqMan Fast Universal Master Mix 2X with a pre-made primer set (Mm01134790_g1) . The relative expression of Ager was normalized to the expression of Ipo8 housekeeping gene. The statistical significance of differences was evaluated by Wilcoxon Signed-Rank Test.Survival probability is defined as the probability of surviving over a specific length of time  by lowering the mouse toward the metal mesh grid, allowing its hind paws to attach to the grid connected to a force transducer and gently pulling backwards with consistent force by the experimenter until the grip was released. The peak force produced during the pull on the bar in grams was recorded by the transducer. Demonstration of the procedure is shown in the Supplementary Movie p < 0.05) was evaluated by  non-parametric ANOVA with Student-Newman–Keuls post-test.All values are presented as mean ± standard error (SEM). The statistical significance of differences , when compared to non-transgenic SJL mice used as a reference, 100%, 472 days, as published , was administered beginning at age 8 weeks until the final stage of the disease, as defined above. Detailed analysis of the survival and longevity span revealed that male mice injected with sRAGE displayed on average a higher probability of surviving for a longer time after the disease onset; 7 Figure , and on 1 Figure . The ons1 Figure . In addiMotor performance scores, as measured by grip strength meter test and hanging cage test Figures , revealeNeuronal loss and increased presence of glial cells (astrocytosis) are hallmarks of ALS pathology in the spinal cord. To determine whether sRAGE treatment affected neuronal loss or astrocytosis, we stained spinal cord samples with cresyl violet, a dye with neuronal cell affinity, and with antibodies to GFAP, a well-established astrocyte maker. We observed significant differences in the number of ventral horn motor neurons Figures  between Our study is the first report of the beneficial effects of treatment with sRAGE, which sequesters RAGE ligands and prevents their interaction with and activation of the cell surface receptor, in delaying the onset and progression of a neurodegenerative disease in the SOD1 transgenic mouse model of ALS. We observed that the onset of ALS, as measured by weight loss and performance scores, was delayed in sRAGE-treated SOD1 mice and that these mice had improved motor functional scores and prolonged life span. Additionally, we observed that these mice consistently, and for longer periods of time from the disease onset, performed better than their control treated-counterparts, manifesting overall better health condition and prolonged life span, as compared to controls. Our study also provides the first molecular evidence that, comparable to that observed in the human, expression of RAGE is increased in the SOD1 transgenic mouse lumbar spinal cord.Ager improves post injury sciatic nerve regeneration in type 1 diabetic mice, at least in part, by switching infiltrating macrophages from pro- to anti-inflammatory signatures, linked most likely to reduction in tissue-damaging inflammation and thereby to improvement in neuronal survival scores at the injury site  of patients has been proposed to serve as a biomarker for disease onset or severity in multiple diseases  mouse model of AD (Deane et al., RAGE antagonists also show some promise for effective treatment in neurodegenerative diseases. Deane and colleagues identified a highly specific small molecule, FPS-ZM1, that blocks Aβ binding to the V-type extracellular domain of RAGE, thus preventing Aβ accumulation and Aβ-induced cellular toxicity in the APP (Our previous investigation on RAGE-ligand expression in human ALS thoracic spinal cord found higher expression of RAGE and its ligands compared to controls (Juranek et al., Riluzole is the first and most commonly used FDA approved drug for ALS treatment. Although an earlier study suggested benefit of Riluzole in the SOD1 transgenic mouse model (Gurney et al., In the light of this report and the absence of any other currently approved treatment, we propose that sRAGE or other forms of RAGE blockade might fill a critical gap in this disorder as a potential single-agent or supplementary candidate for ALS treatment. As AGEs and other RAGE ligands accumulate in the ALS spinal cord, both in the human and in the murine model, we predict that RAGE contributes importantly, at least in part, to the chronic and sustained loss of neurons observed in this disorder. Further studies aimed at extending this first report are required to provide insight into molecular mechanisms of RAGE action in ALS.JJ and GD wrote the manuscript, performed and analyzed experiments. MG, RR, BK, and LK performed experiments. HL analyzed data, AMS wrote the manuscript and analyzed data.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.