The global burden of ischaemic strokes is almost 4-fold greater than haemorrhagic strokes. Current evidence suggests that 25–30% of ischaemic stroke survivors develop immediate or delayed vascular cognitive impairment (VCI) or vascular dementia (VaD). Dementia after stroke injury may encompass all types of cognitive disorders. States of cognitive dysfunction before the index stroke are described under the umbrella of pre-stroke dementia, which may entail vascular changes as well as insidious neurodegenerative processes. Risk factors for cognitive impairment and dementia after stroke are multifactorial including older age, family history, genetic variants, low educational status, vascular comorbidities, prior transient ischaemic attack or recurrent stroke and depressive illness. Neuroimaging determinants of dementia after stroke comprise silent brain infarcts, white matter changes, lacunar infarcts and medial temporal lobe atrophy. Until recently, the neuropathology of dementia after stroke was poorly defined. Most of post-stroke dementia is consistent with VaD involving multiple substrates. Microinfarction, microvascular changes related to blood–brain barrier damage, focal neuronal atrophy and low burden of co-existing neurodegenerative pathology appear key substrates of dementia after stroke injury. The elucidation of mechanisms of dementia after stroke injury will enable establishment of effective strategy for symptomatic relief and prevention. Controlling vascular disease risk factors is essential to reduce the burden of cognitive dysfunction after stroke. This article is part of a Special Issue entitled: Vascular Contributions to Cognitive Impairment and Dementia edited by M. Paul Murphy, Roderick A. Corriveau and Donna M. Wilcock. •Ischaemic injury is common among long-term stroke survivors•About 25% stroke survivors develop dementia with a much greater proportion developing cognitive impairment•Risk factors of dementia after stroke include older age, vascular comorbidities, prior stroke and pre-stroke impairment•Current imaging and pathological studies suggest 70% of dementia after stroke is vascular dementia•Severe white matter changes and medial temporal lobe atrophy as sequelae after ischaemic injury are substrates of dementia•Controlling vascular risk factors and prevention strategies related to lifestyle factors would reduce dementia after stroke This demands enormous resources from healthcare systems Deaths from stroke have declined in high-income countries and many middle- and low-income countries. The key element in this decline is reduced incidence of stroke 2The clinical diagnosis of stroke is usually accurate but the precise type of stroke and exact localization may be less straightforward. Determination of the pathological type of stroke is best achieved by early brain imaging usually computed tomography (CT) or by autopsy confirmation. In western countries, cerebral infarction accounts for approximately 80% of first-time strokes, and parenchymal brain haemorrhage for 20%. In Africa, however, the burden of haemorrhagic strokes are reported to be significantly greater by more than 10% at a ratio of 66:34 ischaemic to haemorrhagic 3Dementia developing after stroke is thought to be a clinical entity to define any type of dementia occurring subsequent to stroke injury irrespective whether it involves vascular, neurodegenerative or mixed processes. It can therefore entail a complex aetiology with varying combinations of large and small vessel disease as well as non-vascular neurodegenerative pathology. The development of dementia after stroke depends on several factors including the location and volume of the stroke, degree of related neuronal damage, presence of pre-existing cognitive impairment or other cerebral pathology. The direct influence of any specific genetic factor(s) is not clear. However, the heritability estimate for all ischaemic strokes was determined to be 38% but varied considerably by subtype with the greatest associated with large vessel (40%) and cardioembolic disease (33%) and lowest with small vessel disease (16%) Cognitive impairment or dementia after stroke is predominantly defined by dementia that occurred within three months after stroke onset. Irrespective, many stroke survivors develop delayed dementia beyond three months or only after recurrent stroke(s). The recognition of cognitive impairment in the acute phase after stroke may offer vital information to the clinician for early cognitive rehabilitation Recent prospective studies suggest stroke survivors may unmask or trigger varied pathologies including those attributed to subcortical VaD, multi-infarct dementia and even strategic infarct dementia The cognitive domains involved in the development of dementia after stroke may also vary depending on stroke characteristics such as stroke type, volume, numbers, location and severity. Regarding the stroke type, patients with ischemic strokes usually have higher survival rates than do those with hemorrhagic strokes, which explains why ischemic strokes lead to psychiatric morbidity more frequently than do hemorrhagic strokes In post-stroke cohorts, the presence of executive syndrome and depression is the predictor of poor long-term survival rather than depression itself Dementia associated with cerebrovascular diseases is clinically diagnosed using the widely accepted DSM IIIR or IV criteria. The apparent “gold standard” for diagnosis has been based on the results of the extensive neuropsychological examination, clinical presentation, and information from a close relative as well as the usefulness of R-CAMCOG Dementia after stroke may incorporate different types of dementias but these there is hardly any pathological confirmation for most of the clinical or prospective studies 4As the risk of death from strokes has declined, the number of stroke survivors with cerebral compromise and cognitive dysfunction has increased. Stroke survivors are at increased risk of cognitive impairment Cognitive impairment after stroke is a frequent but neglected consequence compared to other neurological deficits such as sensory or motor impairment 5In tandem with the age-related increased incidence of stroke itself, older age is the strongest risk factor for VCI and dementia after stroke . This isHypertension is the most common modifiable risk factor for stroke. Elevations in both systolic and diastolic blood pressures are associated with increased risk of stroke and by extension stroke related dementia . In addiStudies from general community populations indicate that VaD is more frequent in males than in females, but most studies suggested no substantial gender difference for the risk of dementia after stroke. Recent findings suggest this is attributable to neuroprotective role of adiponectin because of the relatively steeper decline of serum adiponectin levels associated with ageing in females than in males APOE) gene APOE ԑ4 allele was reported to be associated with greater progression of cognitive decline The risk of dementia is likely more when vascular comorbid conditions occur. Thus, hypertension, atrial fibrillation, diabetes mellitus, myocardial infarction, and congestive heart failure can often co-exist in various proportions NOTCH3 and HTRA1 genes linked to cerebral autosomal dominant (CADASIL) and recessive (CARASIL) disorders. Both common and rare SNPs in the NOTCH3 gene show increased risk of age-related WM changes in hypertensive subjects HTRA1 gene were identified to be associated with hereditary SVD of unknown aetiology COL4A2 gene were identified to be associated with symptomatic small vessel disease, particularly WM disease and deep intracerebral haemorrhage Variants in genes associated with familial small vessel diseases of the brain 6Neuroimaging has been very useful to recognise that a combination or interaction of different types of brain lesion, including neurodegenerative markers, and preexisting underlying processing are players in the development of cognitive decline after clinical stroke. Brain lesion correlates of dementia after stroke include a combination of infarct features , the presence of WM changes , as well as brain atrophy 7Much of the current knowledge of the ischaemic injury cascade comes from experimental studies. The cascade consists of a complex series of events which are highly heterogeneous c and subsequent stimulation of caspase-3 whereas the extrinsic pathway is initiated by the activation of cell surface death receptors, which belong to the tumour necrosis factor superfamily, by Fas ligand, resulting in the stimulation of caspase-8. There is some evidence that ischaemic cell death may also be mediated by autophagy, which is activated during cerebral ischaemia for the bulk removal of damaged neuronal organelles and proteins Experimental studies suggest neuronal death following ischaemic injury is largely attributed to necrosis. However, recent developments indicate that significant cell death occurs by apoptotic as well as hybrid mechanisms (e.g. necroptosis) along an apoptosis–necrosis continuum. While the infarcted core is necrotic be it macro- or microinfarction, within the penumbra caspase-mediated apoptosis is activated although secondary necrosis results from failure to implement the apoptotic signalling pathways fully, as they require energy. Ischaemic injury triggers two general pathways of apoptosis. The intrinsic pathway originates with mitochondrial release of cytochrome Neuroinflammation and immunodepression are also associated with stroke, ageing, and infection. These likely have a detrimental role in cognitive function after stroke Compared to neuroimaging correlates the pathological substrates associated with dementia after stroke or VaD have generally lagged behind. This is particularly true in defining those substrates associated with executive dysfunction. There is selective atrophy (30–40%) of pyramidal cells in layers III and V of the dorsolateral prefrontal cortex compared to the anterior cingulate and orbitofrontal cortices of the frontal lobe in subjects with dementia after stroke, VaD and, of mixed and AD versus normal ageing controls and those who did not have dementia after stroke 8Constant perfusion of the WM by deep penetrating arterioles is essential for functioning of axons and oligodendrocytes. Depending on the duration and severity of ischaemic or oligaemic injury, several features may be readily evident in the WM. These include activated microglia, clasmatodendritic astrocytosis, myelin breakdown, presence of axonal bulbs and degeneration, reactivation and loss of oligodendroglia. During subsequent (chronic) periods of arteriolar changes, perivascular spacing and apoptotic oligodendroglia are seen accompanied by degeneration of myelin and expression of hypoxia markers. WM hyperintensities as seen on brain T2-weighted or FLAIR magnetic resonance imaging (MRI) are associated with varying degrees of cognitive dysfunction in stroke, cerebral small vessel disease and dementia. The pathophysiological mechanisms within the WM accounting for cognitive dysfunction remain unclear. Stroke patients with more severe WM changes have an increased risk of recurrent strokes. Thus, the presence and severity of WM changes seen on MRI may be predictive of post-stroke dementia Recent clinicopathological studies 9Medial temporal lobe and global atrophy are both shown to be associated with dementia after stroke 10Several studies have consistently reported that cerebral silent infarcts detectable with computed tomography or MRI were independently predictive of dementia after stroke 11The risk of dementia after stroke is increased in patients with pre-stroke cognitive decline, with about one-third of patients meeting the criteria for AD and two-thirds meeting the criteria for VaD 12Any measure that reduces or controls vascular disease would be preventative for dementia after stroke Stroke is related to two- to nine fold increase in risks of dementia 13Cognitive impairment after ischaemic stroke injury is common in different populations. Although dementia after stroke is a clinical entity, current neuroimaging and pathological studies suggest that majority of older age-related dementia after stroke can be classed as VaD. Emerging small vessel disease associated genetic traits, severe WM changes and medial temporal lobe atrophy are important features in the development of dementia after stroke injury. The well-established relationship between vascular risk factors and dementia provides a rationale for the implementation of interventions. Control of vascular disease risk and prevention of recurrent strokes are obviously key to reducing the burden of cognitive decline and dementia after stroke.The authors declare no competing interests.Transparency document


Acute myocardial infarction (AMI) is a major cause of death worldwide and affects both global and regional left ventricular function. Regional function can be determined by CMR i.e. wall thickening, and velocity encoded (VE) strain analysis. Aim: The aims of this study were to investigate how regional myocardial wall function, assessed by CMR velocity encoded strain and regional wall thickening, change after acute myocardial infarction and to determine if these variables can be used to differentiate between ischemic, adjacent and remote myocardium after experimentally induced myocardial infarction.Ten pigs underwent baseline CMR study for assessment of wall thickening and VE-strain. Ischemia was then induced for 40-minutes by intracoronary balloon inflation in the left anterior descending coronary artery. During occlusion, 99mTc tetrofosmin was administered intravenously and myocardial perfusion SPECT (MPS) was performed for determination of the ischemic area, followed by a second CMR study. Based on ischemia seen on SPECT, the 17 AHA segments of the left ventricle was divided into 3 different categories . Regional wall function measured by wall thickening and VE strain analysis was determined before and after ischemia.Changes in wall thickening and strain before and after ischemia are shown in Figure Both wall thickening and VE strain decrease significantly in both the ischemic and adjacent myocardium acutely after reperfused coronary occlusion. Differentiation thresholds for ischemic, adjacent and remote myocardium, could be determined. Thresholds will however, have limited applicability due to low sensitivity and specificity.Swedish Research Council, Region of Scania, Swedish Heart-Lung Foundation and the Medical Faculty at Lund University.


Cell Regeneration, Cai and colleagues reported that epithelial sheets derived from human induced pluripotent stem cells (iPSCs) can functionally substitute for tooth germ epithelium to regenerate tooth-like structures, providing an appealing stem cell source for future human tooth regeneration.A major challenge in stem cell-based bioengineering of an implantable human tooth is to identify appropriate sources of postnatal stem cells that are odontogenic competent as the epithelial component due to the lack of enamel epithelial cells in adult teeth. In a recent issue  of  Stem cell-based tissue engineering is a promising approach to replace or repair lost or damaged tissues or even organs in humans. Two major types of stem cells currently used in tissue engineering research and clinical application are embryonic stem cells (ESCs) and adult stem cells. However, ethical concerns in using ESCs and difficulties in isolation, expansion, and differentiation of adult stem cells limit their clinical application. The advent of iPSCs has provided a promising alternative source of stem cells for tissue bioengineering .Whole tooth bioengineering has given hope to dental replacement and regenerative therapy , 3. In tTooth development relies on reciprocal tissue interactions between ectoderm-derived dental epithelium and cranial neural crest-derived mesenchyme . In miceCell Regeneration , Cai and colleagues reported their studies on ameloblastic differentiation capability of integration-free human urine-derived iPSCs (ifhU-iPSCs) [Attempts have been made to identify alternative sources of human postnatal stem cells as the epithelial component for human whole tooth regeneration. It has been demonstrated that keratinocytes isolated from human foreskin and gingival epithelial cells isolated from patients’ gingival tissue are able to differentiate into enamel-secreting ameloblasts when recombined with mouse embryonic molar mesenchyme that possesses odontogenic potential , 20. HowU-iPSCs) . They fiCertainly, identification of novel adult human stem cell sources is not yet the final solution for tooth regeneration. Based on the characteristic features of tooth development, to generate an implantable tooth germ in vitro, one of the cell sources, either epithelial or mesenchymal population, must acquire odontogenic potential to initiate regenerative process. In fact, despite the fact that human iPSC-derived epithelial cells  as well


Aeropyrum pernix and Sulfolobus tokodaii, were determined approximately 15 years ago. In these genome datasets, 47 and 46 tRNA genes were detected, respectively. Among them, 14 and 24 genes, respectively, were predicted to be interrupted tRNA genes. To confirm the actual transcription of these predicted tRNA genes and identify the actual splicing patterns of the predicted interrupted tRNA molecules, RNA samples were prepared from each archaeal species and used to synthesize cDNA molecules with tRNA sequence-specific primers. Comparison of the nucleotide sequences of cDNA clones representing unspliced and spliced forms of interrupted tRNA molecules indicated that some introns were located at positions other than one base 3' from anticodon region and that bulge-helix-bulge structures were detected around the actual splicing sites in each interrupted tRNA molecule. Whole-set analyses of tRNA molecules revealed that the archaeal tRNA splicing mechanism may be essential for efficient splicing of all tRNAs produced from interrupted tRNA genes in these archaea.Based on the genomic sequences for most archaeal species, only one tRNA gene (isodecoder) is predicted for each triplet codon. This observation promotes analysis of a whole set of tRNA molecules and actual splicing patterns of interrupted tRNA in one organism. The entire genomic sequences of two Creanarchaeota,  Many tRNA molecules have been purified from original host organisms, and the respective tRNA nucleotide sequences were determined via direct RNA sequencing. Additionally, tRNA molecules are known to contain many different kinds of modified nucleotides.Mycoplasma carpicolum ..A. perniable arm . The predon stem . We alsotructure A–C 15]..A. pernierformed . These aBy isolating cDNAs representing actual tRNA molecules and identifying the actual splicing events in the archaeal tRNA system, we have contributed important and useful information on expression and processing of tRNA genes.


Candida species are the most frequently found fungal pathogens causing nosocomial disease in a hospital setting. Such species must be correctly identified to ensure that appropriate control measures are taken and that suitable treatment is given for each species. Candida albicans is causing most fungal disease burden worldwide; the challenge lies in differentiating it from emerging atypical, minor and related species such as Candida dubliniensis and Candida africana. The purpose of this study was to compare identification based on MALDI-TOF MS to standard identification systems using a set of nosocomial isolates.HWP1).Eleven nosocomial samples were collected from 6 third-level hospitals in Bogotá, Colombia. All the samples were identified by combining MALDI-TOF MS with morphological characters, carbohydrate assimilation and molecular markers  and cluster analysis, differences being revealed between Candida albicans, Candida africana, Candida dubliniensis reference spectra and two clinical isolate groups which clustered according to the clinical setting, one of them being clearly related to C. albicans.The present work describes the first collection of atypical Colombian Candida albicans-related and atypical C. albicans species, thereby overcoming conventional identification methods. This is the first report of hospital-obtained isolates of this type in Colombia; the approach followed might be useful for gathering knowledge regarding local epidemiology which could, in turn, have an impact on clinical management. The findings highlight the complexity of distinguishing between typical and atypical Candida albicans isolates in hospitals.This study highlights the importance of using MALDI-TOF MS in combination with morphology, substrate assimilation and molecular markers for characterising  Candida albicans is the pathogenic fungus most commonly found in the general population; its ease of transmission in hospitals causes high comorbidity rates  rCO_B isolates had a common hospital origin and more diverse clinical settings, lacking antifungal prophylactic guidelines. This factor could lead to less stressful conditions for nosocomial isolates and less selective pressure; the latter has been considered by Maubon et al., (2014)  as beingC. albicans isolates require further molecular and susceptibility studies for understanding their ability to adapt (this includes phenotypic plasticity) and their potential for harming a vulnerable host.The clinical and epidemiological aspects of infection are relevant, since the pathological implications of these atypical


The prodigious capacity of our brain to process information relies on efficient neural coding strategies. In engineered systems, bandwidth is often increased through multiplexing - multiple signals are simultaneously, yet independently, transmitted through a single communication channel. We have proposed previously that neural systems might implement the same sort of solution . Here, wa priori knowledge of which signal evokes which spikes) whereas treating all spikes as equivalent compromised STA measurement, especially for the fast signal (meaning that one needs to subdivide spikes according to their level of synchrony). Beyond simply measuring STAs, Figure To test our hypothesis, we built a feed-forward neural network comprising Morris-Lecar (ML) model neurons. All neurons received a common input constructed from two distinct signals, slow and fast, plus uncorrelated fast noise. According to our hypothesis, slow and fast signals are independently encoded by different types of spikes; in other words, differentially correlated output spikes, namely, asynchronous (Async) and synchronous (Sync), enable encoding of slow and fast signals, respectively. To assess the feasibility of the multiplexed coding scheme, recorded spikes were classified into two independent classes based on the peristimulus time histogram (PSTH) calculated from the entire set of neurons. Spikes whose instantaneous rates exceeded a threshold were designated "Sync" and all others were designated "Async". The spike triggered average (STA) was calculated for slow and fast signals using Sync and Async spikes, resulting in four different STAs. The Async-slow and Sync-fast STAs were clearly structured whereas the other two were not Figure . Using tOur results demonstrate the feasibility of multiplexed coding using synchronous and asynchronous spiking. Decoding the two signals requires that spikes are distinguished by their cross-correlation; this is difficult to do with experimental data in which only a subset of neurons are recorded, but it is straightforward for the brain using downstream neurons that operate as coincidence detectors or integrators that respond preferentially depending on input correlation.


Habitat loss and fragmentation are leading causes of species extinctions in terrestrial, aquatic and marine systems. Along coastlines, natural habitats support high biodiversity and valuable ecosystem services but are often replaced with engineered structures for coastal protection or erosion control. We coupled high-resolution shoreline condition data with an eleven-year time series of fish community structure to examine how coastal protection structures impact community stability. Our analyses revealed that the most stable fish communities were nearest natural shorelines. Structurally complex engineered shorelines appeared to promote greater stability than simpler alternatives as communities nearest vertical walls, which are among the most prevalent structures, were most dissimilar from natural shorelines and had the lowest stability. We conclude that conserving and restoring natural habitats is essential for promoting ecological stability. However, in scenarios when natural habitats are not viable, engineered landscapes designed to mimic the complexity of natural habitats may provide similar ecological functions. Coastal habitats host diverse ecological communities and provide numerous ecosystem services that affect the health, security and quality of life of human societies ,2. The dThe concepts of stability and resilience have been central foci of both fundamental ecology and applied conservation for at least the past half-century –14. StabThe utilization of engineered coastal structures such as vertical walls and revetments directly replaces natural shoreline habitats, disrupts land-water exchange, and alters the biophysical environment , potentially indirectly harming other natural habitats ,21,22. OAlthough the societal and ecological costs of coastal habitat degradation are becoming increasingly recognized –25, coasThis study involves the analysis of data resulting from routine fisheries research and monitoring efforts by the State of Alabama Department of Conservation and Natural Resources—Marine Resources Division (ADCNR-MRD). ADCNR-MRD research and monitoring is regulated and permitted by the State of Alabama. ADCNR-MRD’s standard protocol for sampling vertebrate fishes involves the live release of specimens when possible. This study did not involve endangered or protected species. The data analyzed and interpreted in this study were collected in Mobile Bay, Alabama USA .2 of bay water surrounded by 235 km of shoreline  . The GSAEleven years of monthly data on coastal fish communities (2001–2011) were acquired from a fishery-independent state survey designed to monitor juvenile and adult finfish populations. The survey was initiated in 2001 with a randomly stratified sampling scheme throughout five zones in the northern and southern regions the bay  across shoreline types using permutational multivariate analysis of variance (PERMANOVA) and non-metric multidimensional scaling (nMDS) analysis on Bray-Curtis distances . To quanAnalysis of the Euclidean distance in ordination space between mean annual fish community structure observed in all pairs of years revealed that temporal variability of fish communities was related to shoreline condition . CommuniTo determine whether these stability differences between shoreline conditions were present at sub- and super-annual periods, we computed the global wavelet power spectrum, which measures the average temporal variability at each period from 2001 to 2011. For average abundance, communities associated with natural shorelines had much lower temporal variance  than that of the engineered shorelines at periods ranging from 2–40 months . For speWe found that natural landscapes support more stable fish communities than engineered landscapes. However, not all engineered landscapes performed identically. For instance, the stability of coastal fish communities was significantly higher near engineered shorelines characterized by rubble and riprap revetments than vertical walls or vertical walls with riprap. This finding suggests that structural complexity can in some instances reduce the negative effect of engineered structures on community stability. Natural habitats such as saltmarsh, oyster reef and submerged aquatic vegetation are structurally complex and widely recognized for providing essential habitat and nursery grounds for a variety of coastal species –34. On tBy analyzing community similarity and variability at annual and monthly intervals, we assessed typical fluctuations or trends of stability and evaluated the potential impacts of discrete events of disturbance. For both the sequential and overall annual time series analyses, communities associated with natural shorelines exhibited higher community similarity and fluctuated less than all engineered shoreline conditions. The global wavelet power analyses indicated that the high resilience of natural landscapes, which was observed at annual timescales, also applies at shorter time periods. During the 11 years that were examined in our study, the Alabama Gulf coast was impacted by several hurricanes and tropical storms including Allison in 2001, Ivan in 2004, Dennis, Katrina and Rita in 2005 and Ike in 2008. The Gulf of Mexico also experienced a massive oil spill following the explosion of the Deepwater Horizon drilling rig in 2010. However, studies of tidal marsh creeks following Ivan and seagrass meadows following Katrina found very little impact of the hurricanes on coastal habitats ,37. The Although engineered shorelines that mimic the complex structure of natural coastal habitats can partially restore community stability at local scales, preserving natural habitats may be important for community stability at both local and regional scales by “spilling over” via dispersal. Indeed, in fluctuating and interconnected metacommunities experiencing different environmental conditions , connectivity can have a large impact on community stability across scales ,40. In tThe legacy and extraordinary degree of shoreline alteration in Mobile Bay, like many other coastal systems, dates back far longer than comprehensive ecological monitoring, making it quite challenging to understand how current fish communities adjacent to different shoreline types actually compare to a natural coastal community. However, the emergence of landscape ecology and the availability of longer term data series on ecosystem change have greatly improved our ability to understand how human activities have transformed the structure and function of natural landscapes –46. For S1 File(DOCX)Click here for additional data file.S2 File(DOCX)Click here for additional data file.


Smoking during pregnancy increases the risks of many pregnancy-related complications. MiQuit is a tailored, self-help, text-message intervention developed to help pregnant smokers to stop. A pilot RCT investigated feasibility of recruitment of women attending hospital antenatal clinics by using NIHR Clinical Research Network (CRN) research midwives (RMs); findings will be used to plan a future definitive trial to investigate MiQuit efficacy. We aimed to describe the facilitators and barriers to trial recruitment that RMs perceived, and relate these to key recruitment processes.We conducted 14 semi-structured telephone interviews with RMs from 13 of 15 pilot trial recruiting centres. All interviewees had been involved in local study set-up, participant recruitment and follow-up. Data were transcribed verbatim, transcripts were coded simultaneously and inductive thematic analysis was used to analyse data.Emergent findings suggest a number of pertinent themes, most notably with participant identification and screening. For example, experienced RMs generally felt that a screening questionnaire intended to be given to all women attending antenatal clinics, and therefore prevent smokers from feeling unfairly targeted, was ineffective when used in the proposed manner and could cause greater discomfort on both sides. Instead, RMs indicated that they preferred to pre-identify smokers and discreetly approach them.Qualitative exploration of research staff views can help maximise the value of pilot work, identifying key aspects of trial design and conduct that may affect recruitment. We will discuss how these findings can assist planning a definitive trial, highlighting issues which may be generalisable to other studies.


Copy number variations (CNVs) consist of duplications or deletions of chromosomal regions ranging from a few hundred to more than a million bases in size, and are likely to play a role in phenotypic diversity and evolution. Recent advances in the identification and mapping of CNVs among normal individuals and in model systems, using bioinformatics and hybridization-based methods, are beginning to shed light on the functional importance of CNVs.Most CNVs are harmless; however, some are associated with human diseases including neurodevelopmental disorders. Hundreds of CNVs have been linked to neurological phenotypes, including autism, schizophrenia, and bipolar disorder. Some CNVs, such as duplications involving VIPR2 on 7q36.3 and deletions of NRXN1 on 2p16.3 have been identified in autism, schizophrenia, and bipolar disorder [Recently, model systems for human genetic disorders have emerged based on the use of human pluripotent stem cells (hPSCs). These approaches include human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). These models offer unique advantages for the study of developmental biology. hPSCs can be differentiated into various types of somatic cells including neurons to examine molecular and cellular mechanisms involved in disease causation. Differentiated hiPSCs have enabled the development of human in vitro model systems that capture the inherent pathologies based on the genetic background of the source material. These stem cell lines derived from patients with various diseases offer unique advantages for the study of inherited neurological disorders. Such patient-specific hiPSCs allow examination of underlying genetic factors in human organ systems that affect neural development.MRI scans and post-mortem studies of autism spectrum disorders suggest abnormal brain development due to initial brain overgrowth followed by premature growth arrest . DuplicaSchizophrenia is a complex genetic neurodevelopmental disorder that carries increased CNV burden. Using iPSC technology, Brennand and coworkers showed that hiPSC neurons from schizophrenia patients exhibit diminished neuronal connectivity and reduced neurite outgrowth and glutamate receptor expression . In addiNovel genome engineering approaches using ZFNs, TALENs, and CRISPRs have been applied to study specific functions in hPSCs. These tools provide the opportunity for correction of disease-causing mutations in patient-derived hiPSC models. Importantly, genetic and epigenetic instabilities, including enhanced de novo CNV induction have been observed during long-term propagation of hPSCs . These cStem cell technology using patient-derived hiPSCs with well-defined CNVs recapitulates the developmental programs that guide formation or malformation of brain region-specific neurons. The application of genomeediting tools to studies of hiPSCs will give important insights into the genome structural changes that underlie fundamental disease processes at the cellular level. These combined approaches are a boost to developmental neuroscience that could be transformative in the years ahead.


Nicotiana species in the graft union zone, within a tissue culture system. This has led to the formation of alloploid cells that, under laboratory conditions, gave rise to a novel, alloploid Nicotiana species, indicating that natural grafts may play a role in plant speciation, under certain circumstances.Grafting, an old plant propagation practice, is still widely used with fruit trees and in recent decades also with vegetables. Taxonomic proximity is a general prerequisite for successful graft-take and long-term survival of the grafted, composite plant. However, the mechanisms underlying interspecific graft incompatibility are as yet insufficiently understood. Hormonal signals, auxin in particular, are believed to play an important role in the wound healing and vascular regeneration within the graft union zone. Incomplete and convoluted vascular connections impede the vital upward and downward whole plant transfer routes. Long-distance protein, mRNA and small RNA graft-transmissible signals currently emerge as novel mechanisms which regulate nutritional and developmental root/top relations and may play a pivotal role in grafting physiology. Grafting also has significant pathogenic projections. On one hand, stock to scion mechanical contact enables the spread of diseases, even without a complete graft union. But, on the other hand, grafting onto resistant rootstocks serves as a principal tool in the management of fruit tree plagues and vegetable soil-borne diseases. The ‘graft hybrid’ historic controversy has not yet been resolved. Recent evidence suggests that epigenetic modification of DNA-methylation patterns may account for certain graft-transformation phenomena. Root grafting is a wide spread natural phenomenon; both intraspecific and interspecific root grafts have been recorded. Root grafts have an evolutionary role in the survival of storm-hit forest stands as well as in the spread of devastating diseases. A more fundamental evolutionary role is hinted by recent findings that demonstrate plastid and nuclear genome transfer between distinct  Cucurbitae and Solanaceae species. Thus, the majority of recent grafting research concerns physiological and pathological aspects of vegetable grafting. Grafting also plays an important role in various types of physiological investigations . A broad range of classical grafting techniques can be found in igations , in partigations . An overigations . The hisigations . Undoubtresearch  has openCapsicum annuum L.), old-stage homografts (58 days old plants) had slower graft-take than young-stage (34 days) homografts. Old-stage grafts had a lower percentage of xylem connections and seemed to suffer from drought stress  are presumably always compatible. In heterografts, broadly speaking, intraspecific grafts  are nearly always compatible, interspecific grafts (= rootstock and scion belonging to different species of the same genus) are usually compatible, intrafamilial grafts are rarely compatible, and interfamilial grafts are essentially always incompatible . Examinat stress . The timrequired  but Yin ts e.g., .Graft incompatibility may be defined as failure to form a successful graft union. Yet, despite innumerous follow-ups of graftage in various types of plants, the reasons for graft incompatibility are still vague. Initial healing of the graft union does not in itself ensure long-term compatibility. In cucurbits, apparently successful grafts proved incompatible 25 days after grafting . In certSolanaceae, reciprocal grafts of tomato (Solanum lycopersicum L.) and pepper were considered severely incompatible, whereas tomato and eggplant (Solanum melongena L.) only moderately so, in comparison with compatible tomato homografts (Cucumis melo L.) onto 22 Cucurbitae rootstocks (Cucurbitae species (Castanea) species  onto dwarfing rootstocks  scions and quince (Cydonia oblonga Mill.) rootstocks. Prunasin, a cyanogenic glucoside, rises from the quince rootstock into the pear scion, where it is catabolized enzymatically by pear glycosidase to liberate cyanide at the graft interface. Cyanide ‘poisons’ the graft union tissues, causing cellular necrosis at the graft interface, leading to severe incompatibility (Prunus persica L. Batsch) grafted on myrobalan plum (Prunus cerasifera L. Ehrh) suggested a more general poisoning mechanism, due to progressive impairment of the phloem carbohydrate transport and accumulation of starch above the graft union , which presumably involves some kind of mobile RNA, was one of the pioneering findings in this area , in a distant organ , does not in itself prove its vascular transport, since this compound could also be synthesized in the presumably recipient organ. Therefore, grafting of genetically distinct mutants/transgenes is still being used as a principal means to distinguish between the site of biogenesis and the recipient organs . Roots oagnitude . FurtherSolanum tuberosum L.) tuberization by the graft transmissible miR156 is another recently studied case (Gibberellic Acid Insensitive (GAI) mRNA, including the anticipated changes in plant phenotype  through a variety of tracks, including natural, underground root grafts. This avenue of disease transmission has been underestimated , just asSantalales) and dodder (Cuscuta sp.) transmit pathogens very efficiently, even among intergeneric and interfamilial plant species (Parasitic plants such as mistletoes (= parasitic plants in the order  species . Dodder  species .Persea americana Mill.) transferred a citrus viroid to avocado; the grafted citrus budwood remained viable for almost a year without forming a true graft union (Catharanthus sp.) seems to be unique in its ability to form distant interspecific grafts. The apple proliferation phytoplasma disease was transmitted from infected apple scions to periwinkle rootstocks  rootstocks to overcome the Phylloxera grapevine plague (Citrus tristeza virus (Closterovirus) and other virus-like diseases exploited every possible graft combination  had PGIP activity. PGIP was detected in xylem exudates of untransformed grapevine scions grafted onto transgenic rootstocks expressing pPGIP, indicating that the PGIP protein is graft–transmissible. Such scions revealed improved tolerance toward grapevine diseases caused by Xylella fastidiosa and Botrytis cinerea  also seem to play a role in rootstock-induced disease resistance. Polygalacturonase-inhibiting proteins (PGIPs) are plant cell proteins that specifically inhibit the cell wall degrading endo-polygalacturonases of plant pathogens . Leaf ex cinerea . Followi cinerea . This idFigure 2).Of all grafting issues, the least understood and most controversial is the ‘graft hybrid’ concept. According to this concept grafting may involve stock to scion transfer of genetic material (= graft transformation), leading to heritable changes in the scion. The scion which has acquired certain heritable traits from the rootstock is regarded as a ‘graft hybrid.’ However, graft transformations occur only under ‘Mentor grafting’ conditions, which presumably enforce the transfer of genetic material from stock to scion  are removed throughout the experiment. Stock fruits are also removed in attempt to maximize flow of substances from the stock to the scion . InteresThe graft hybrid concept, which was developed and demonstrated by the Soviet horticulturist Solanaceae, in particular pepper (Within the ‘graft hybrid’ supportive evidence some specific characteristics can be defined. (a) The frequency of the appearance of variant plants is highly variable, sometimes below 1% . Thus, tr pepper , which ir pepper  has beenr pepper  but no fIn their review, Epigenetics refers to reversible heritable changes in genome function that occur without a change in the DNA sequence and may have morphological, physiological, and ecological consequences . ChangesSolanaceae . Root grafting could be induced artificially between potted l months . Naturall months . Althougl months , it may Assessment of the evolutionary significance of grafting involves discussion at two levels: its benefits for plant survival and its potential role in the formation of new species. The plausible ecological benefits of root grafting have been discussed by Addressing the significance of grafting as a potential evolutionary mechanism must take into account the ancient belief that grafting may give rise to new plant species . The ‘grNicotiana species (Recent research has brought us much closer to a potentially real evolutionary grafting mechanism. In their early report  species . Interes“… do not lend support to the tenet of Lysenkoism that ‘graft hybridization’ would be analogous to sexual hybridization. Instead, our finding that gene transfer is restricted to the contact zone between scion and stock indicates that the changes can become heritable only via lateral shoot formation from the graft site.”Nicotiana species was thus obtained, tentatively named Nicotiana tabauca. Still, natural evolutionary formation of new species via this pathway would depend upon natural graft formation and emergence of adventitious lateral shoots, as previously discussed.However, in a more recent report  Bock andNicotiana under tissue culture conditions require further confirmation with other plants in natural settings (The hypothetical possibility of cell fusion graft hybrids has already been mentioned by settings . It is dArabidopsis model has opened the way for meticulous, in-depth grafting research. The currently available molecular tools are expected to advance our understanding and eventually resolve the long standing grafting mysteries.While the technology of grafting has advanced tremendously, the long-term survival of grafted, composite plants is still somewhat unpredictable. The complexity of stock/scion interactions attains a new dimension when pathogenic agents enter the scenery. More enigmatic are the graft hybrid conundrum and the broader evolutionary significance of grafting. The extensive horticultural grafting research has not usually addressed these basic questions. The recently adopted 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.


This is a case report of 22-year-old girl admitted with abdominal distension, vomiting, and chronic constipation since birth. Abdomen was distended, and perineal examination revealed imperforate anus with vestibular fistula (ARM). So far worldwide very few cases have been reported about anorectal malformation presenting in adulthood, and thus extremely little data is available in the literature about an ideal management of anorectal malformation in adults. In our case in the treatment instead of conventional procedure of posterior sagittal anorectoplasty (PSARP) anal transposition was done and till two years after the definitive treatment during follow-up patient has been doing well with Kelly's score of six. Our experience suggests that anal transposition provides satisfactory outcome in adults presenting late with anorectal malformation. Though anorectal malformation (ARM) is pediatric problem, in India because of poverty and ignorance one may encounter it in adults also. Posterior sagittal anorectoplasty (PSARP) is now accepted as a standard treatment in children, but there is very little literature guiding an ideal treatment of anorectal malformation in adults. Here we present a case of an adult female who was neglected since childhood and presented with congenital low type of anorectal malformation, we performed anal transposition which gave her a good outcome.22-year-old female presented with features suggestive of large bowel obstruction, chronic constipation since birth and regular passage of small quantity of stool through an opening in the perineum. Perineal examination revealed no separate anal opening , dry stoTransverse diameter inside the fistulous opening appeared very large on P/R exam and stool gave a firm feel to finger. X-ray abdomen showed loaded colon. She had no other congenital anomalies. In the first setting after primary resuscitation an emergency laparotomy was carried out and proximal diverting transverse loop colostomy was done to relieve the obstruction. After this under anesthesia stepwise instrumental evacuation of stool was performed through fistulous opening as hard and enormous quantity did not yield to conservative treatment, through either stoma or distal fistula. Few days later distal cologram showed decompressed distal colon and finally patient was planned for definitive procedure.Under spinal anaesthesia patient was put in lithotomy position, thorough examination was done, and previous clinical findings were confirmed. Tractions sutures applied to expose the operative area , Inj. AdDiagrammatic representation of the procedure has been shown sequentially in Figures Skin sutures were removed after 12 days and 2 weeks after the procedure sequential anal dilatation started, patient was continent. For 2 months along with anal dilator she was maintained on stool softener and high fibre diet. After 2 months stool softener use was tapered. Three months later distal loop cologram showed patent passage, so temporary colostomy was closed. Her anal manometry study was satisfactory. She had voluntary defecation without soling or constipation. She was advised to strictly follow dietary modification onwards. We used Kelly's score to assess her physiologic function during follow-up. At the end of 1 year she was continent without soiling or constipation. She could differentiate between feces and flatus and had strong effective squeeze, with Kelly's overall score of six. Though no objective evaluation was done, she was satisfied with perineal cosmetic outcome.The most common anorectal malformation defect in females is imperforate anus with vestibular fistula. In females this anovestibular fistula is a low type of disease and it opens near the vagina at the posterior fourchette and is directed posteriorly and upward with adhesion to posterior vaginal wall , 2. In aWe feel anal transposition provides satisfactory results in adult patients and also those presenting with anorectal malformation, it provides clear recognition of sphincteric complex with good cosmetic and functional outcome, and result is comparable with PSARP done for children.


The inflammatory response of sepsis results in organ dysfunction, including myocardial dysfunction. Myocardial dysfunction is particularly important in patients with severe septic shock who progress to a hypodynamic pre-terminal phase. Multiple aspects of this septic inflammatory response contribute to the pathogenesis of decreased ventricular contractility. Inflammatory cytokines released by inflammatory cells contribute as does nitric oxide released by vascular endothelium and by cardiomyocytes. Endotoxins and other pathogen molecules induce an intramyocardial inflammatory response by binding Toll-like receptors on cardiomyocytes that then signal via NF-κB. These processes alter cardiomyocyte depolarization and, therefore, contractility. The particular role of the cardiomyocyte sodium current has not been characterized. Now new information suggests that the septic inflammatory response impairs normal depolarization by altering the cardiomyocyte sodium current. This results in decreased ventricular contractility. This is important because new targets for therapeutic intervention can be considered and new approaches to evaluation of this problem can be contemplated. Critical Care, Koesters and colleagues [In the previous issue of lleagues  contribulleagues . Vasodillleagues . It coulThe septic inflammatory response involves a surprising number of intersecting pathways, many of which contribute to ventricular dysfunction. Pathogen-associated molecular patterns (PAMPs) released by infecting organisms bind innate immune receptors on inflammatory cells  but also bind innate immune receptors in the heart  and withCritical Care Koesters, Engisch and Rich investigate the role of cardiomyocyte sodium current [Alteration of intracellular calcium flux is a fundamental later step in the mechanistic pathway to decreased cardiomyocyte and ventricular contractility. In the previous issue of  current . AlteratTo determine whether other channel currents could have contributed, additional characteristics of the septic action potential were measured. There was no significant change in resting potential, a property influenced by non-voltage gated K channels. Membrane resistance and action potential duration decreased only slightly in septic action potentials, suggesting a limited role, at best, for voltage gated K and Ca channels. Inhibition of the L-type Ca current reduced contractility, as expected.Thus, these novel results raise the possibility that sepsis-induced decreases in sodium channel flux may contribute to ventricular dysfunction during sepsis. This conjecture is based on similarity between sepsis-induced and tetrodotoxin-induced changes and therefore is not mechanistic proof of a role of the sodium channel. Nevertheless, moving the focus of investigation towards intracellular signaling expands our knowledge of potential mechanistic pathways contributing to myocardial dysfunction of sepsis. This is important because new targets for therapeutic intervention can be considered and new approaches to evaluation of this problem can be contemplated, possibly even based on surface measurements of electrical properties of the heart.ICAM: Intracellular adhesion molecule; NF-κB: Nuclear factor kappa B; NO: Nitric oxide; PAMP: Pathogen-associated molecular pattern.The author declares that he has no competing interests.


Microvascular disease results in reduced skin blood flow (SkBF) and an increased risk of poor healing, ulceration and amputation, particularly in the lower extremity. Regular exercise is known to produce significant cardiovascular benefits and improved functional outcomes in people with chronic disease. However, it is unknown if these benefits also translate into improvements in SkBF. The purpose of this study was to evaluate the efficacy of exercise training on altering SkBF in adults by systematic review and meta-analysis.Relevant databases were searched to July 2014 for controlled trials evaluating the effect of exercise training interventions versus a non-exercise control on SkBF in adults.Eight studies met the inclusion criteria for this review. Individual studies employing an exercise intervention tended to have small sample sizes, with six of the eight studies showing a benefit of exercise but only three reaching statistical significance. Subsequent meta-analysis demonstrated aerobic exercise had a statistically significant effect on improving SkBF .To date, individual studies employing an exercise intervention have lacked sufficient power to detect clinically relevant benefits to SkBF, partially due to limited sample size. In primarily healthy previously sedentary adult cohorts, pooled analysis revealed a clear benefit of regular aerobic exercise on improving SkBF. Regular aerobic exercise provides a cost-effective option for improving SkBF in adults and may be of benefit to those with cardiovascular disease and metabolic disorders such as diabetes.


Graft ischemia results in inhibition of Na+–K+ ATPase, inhibition of K+ ATP channels, drop of intracellular K+, and the absence of flow favors cell membrane depolarization (+ ATP channels would be associated with increased NADPH oxidase (NOX2) activity and increased production of reactive oxygen species (ROS)  . Decreaspro IL18 . Both IL++-sensitive K+ channels during lung graft ischemia would be expected to provide protection through antagonizing membrane depolarization , which would attenuate ROS production, leading to abortion of inflammasomes priming and activation, and accordingly the release of pro-inflammatory cytokines.Accordingly, the enhancement of Caex vivo lung perfusion , which correlated with decreased incidence of primary graft dysfunction and chronic lung allograft dysfunction after transplantation (++-activated K+ channels (The Toronto team of lung transplantation has achieved significant inhibition of cytokines production within the lung graft through gene therapy during antation . Howeverantation . HO-1 cachannels .++-activated K+ channels during lung graft ischemia. Accordingly, further studies should be conducted to investigate the actual status of these channels during lung graft ischemia prior to transplantation. In addition, pharmacological activation of these channels could be a good target to protect the lung graft during transplantation, with corresponding improvement of the clinical outcome.These findings highlight the possible protective role of the enhancement of CaThe 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.


This project is set out to: i) Establish a collection of astrocytoma cell lines generated from Saudi patients, ii) Select drug resistant brain tumour CSCs, iii) Characterize drug resistant brain tumour CSCs individually, and iv) Deduce common features for drug resistant brain tumour CSCs.The Cancer Stem Cells (CSCs) hypothesis proposes that malignant brain tumours are organized into aberrant cell hierarchies where a subset of parent CSCs replicate asymmetrically and unlimitedly to produce differentiated daughter cell ,2. Thesein vivo tumourgenic assays.A range of methodologies will be applied including, tumour cell line derivation methods, the clonogenic selection assay, cytogenetics profiles, neuronal cancer stem cells characterisation assays and Optimisation of methods used to establish astrocytoma cell lines generated from Saudi patients was completed. To date two novel primary astrocytoma cell lines have been derived. Full clinical data related to the cell lines was retrieved. Currently, optimisation of the clonogenic selection assay and immunostaining for cancer stem cells markers are under progress.Successful retrieval of primary astrocytoma cell lines was possible to accomplish. Important clinical information data relevant to the cell lines were obtained providing clear identification reference. Further work is required for the characterization of the drug resistant cancer stem cell component within these cell lines. Once completed, this research will assist in understanding some of the molecular mechanisms relevant to drug resistance, especially for patients in the Kingdom of Saudi Arabia.


Cumulative studies during the past 30 years have established the correlation between opioid abuse and human immunodeficiency virus (HIV) infection. Further studies also demonstrate that opioid addiction is associated with faster progression to AIDS in patients. Recently, it was revealed that disruption of gut homeostasis and subsequent microbial translocation play important roles in pathological activation of the immune system during HIV infection and contributes to accelerated disease progression. Similarly, opioids have been shown to modulate gut immunity and induce gut bacterial translocation. This review will explore the mechanisms by which opioids accelerate HIV disease progression by disrupting gut homeostasis. Better understanding of these mechanisms will facilitate the search for new therapeutic interventions to treat HIV infection especially in opioid abusing population. Substantial epidemiologic studies report a greater risk of human immunodeficiency virus (HIV) infection in opioid addicts . AlthougRecent studies focusing on the gastrointestinal (GI) immune system illustrate that the gut associated lymphoid tissue (GALT) is the primary target of HIV during all stages of infection  and the The GI tract is considered the largest immunologic organ in the body and plays an important role in maintaining host immune homeostasis. Maintaining gut homeostasis requires balanced interactions among specialized epithelial cells, specialized lymphoid tissues, and commensal bacteria .Figure 1; The intestinal epithelium is the first line of defense in the gut luminal environment. The well-organized intestinal epithelial cells not only provide physical barrier preventing potential pathogen or antigen invasion, but also is integral for nutrient support and water transport thereby maintaining homeostasis of the whole organism . The smaThe major function of the colon is reabsorption of water and any remaining soluble nutrients from the food. The lack of villi in the colon results in a much smaller surface area compared to the small intestine. Organizationally, the epithelial stem cells are located in the lower parts of gut crypts and the differentiated cells migrate to the apical position and populate the colonic epithelial surface .To avoid para-cellular leak of bacteria from gut lumen, the intercellular junctions known as tight junctions join adjacent cells together by extending from the cytoskeleton proteins of one cell into the extracellular matrix, finally joining with the tight junction proteins of another cell . Tight jFigure 2). These cells can modulate the intestinal microenvironment by secreting antibodies, cytokines, or chemokines as well as mediate interaction with the enteric nervous system (ENS).Different types of immune cells are distributed in GALT like PPs and isolated lymphoid follicles, mesenteric lymph node (MLN), and the lamina propria. The major immune cell population within gut tissues includes macrophages, dendritic cells, mucosal mast cells, neutrophilic granulocyte, eosinophilic granulocyte, T lymphocytes, and plasma cells -17A, IL-17F, and IL-22  includes dendritic cells, macrophages, and neutrophils. These cells recognize antigens from the gut lumen and present them to adaptive lymphocytes (T cells and B cells) to initiate the immune responses. Various cytokines and chemokines produced by these APCs are also involved in the regulation of intestinal inflammation . Most land IL-22 . The rolnization .Taken together, both adaptive and innate immune cells are collectively important for maintaining homeostasis within the GI tract. Disruption of any cellular component may potentially predispose hosts to infections or initiate an abnormal immune response that accelerates disease progression.Salmonella due to impaired IgA production and diminished T cell response  μ-receptors, (ii) δ-receptors, (iii) κ-receptors, and (iv) non-classical opioid receptors . OriginaFigure 3), opioids exert both pharmacological and adverse effects in the gut, including alleviating abdominal pain, modulating GI motility, and suppressing intestinal immune functions.By activating these opioid receptors in intestinal tissues  in the infected individuals. All these studies consistently showed that opioid abusers experience more severe neurocognitive defects including HIV-associated dementia , 2006. FFigure 5). As previously described, both HIV/SIV infection and morphine treatment can induce the intestinal epithelial barrier damage. Indeed, humans infected with HIV who use heroin have been shown to have greater amounts of LPS in their serum compared with non-users  and HIV-1 susceptibility (CCR5 and α4β7 integrin) were also increased in morphine-treated animals . Recent Cumulative studies focusing on gut homeostasis help us recognize the central role of gut mucosal tissues in the pathogenesis of AIDS following HIV infection. Virus infection can induce compromised gut epithelial barrier function, abnormal immune activation within the gut tissues, and the dysbiosis in the gut microbiome, which in turn facilitates HIV replication and the disease progression.Additionally, more recent studies demonstrate that opioid abuse can disrupt gut homeostasis in the HIV-infected individuals. The severe disruption of gut homeostasis observed with opioids and models of HIV infection supports what is observed in the clinical studies as HIV patients who inject opioids have higher levels of bacterial translocation than patients who do not use opioids, which is likely why HIV patients who abuse opioids have more severe pathogenesis of HIV and develop AIDS much more quickly than non-users. However, the molecular mechanisms underlying the interactions of opioid abuse and HIV-induced enterotoxicity are still not well understood. Better understanding of these mechanisms may allow for therapeutic interventions to prevent or slow the pathogenesis in the gut following HIV infection, which will finally lead to more powerful strategy to control HIV disease progression especially in the opioid using and abusing population.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.


Bilateral vocal fold immobility may result from bilateral recurrent laryngeal nerve paralysis or physiologic insults to the airway such as glottic scars. The progression of mucosal injury to granulation tissue, and then posterior glottis stenosis, is an accepted theory but has not been photodocumented. This paper presents serial images from common postintubation injury to less common posterior glottic stenosis with interarytenoid synechia. Various pathologic conditions may be observed by the otolaryngologist following endotracheal intubation, including edema, ulcers, cartilage exposure, granulation tissue formation, or glottic stenosis. Management of the most common injuries is often conservative as granulation tissue and ulceration typically resolve spontaneously. The patients with progression to granulation formation are at risk of interarytenoid adhesion formation or early posterior glottic stenosis (PSG) and, therefore, should be closely monitored. While the majority of patients show some laryngeal injury after two days , less thIntubation trauma to the posterior commissure mucosa is theorized to cause ulceration, granulation, and then scar formation, which can fix the vocal cords , 2, 4–7.While numerous articles have described treatment methods for various stages of PGS based upon this theory, none has documented the progression from intubation injury to early posterior glottic stenosis , 9. We pInstitutional review board (IRB) exemption was obtained. This retrospective review of two cases demonstrates early posterior glottic stenosis formation after intubation. Patients complained of progressive voice complaints and exhibited reduced vocal fold mobility. The photomicrographs shown here document the progressive changes over two months (patient 1) and 6 months (patient 2) after acute endotracheal tube injury. Patient 1. A 23-year-old male with a history of traumatic multilimb amputation injury was emergently intubated. He presented with symptoms of acute dysphonia, breathy voice, and vocal fatigue following extubation.Early endoscopic examination of the patient after his hospitalization revealed granulation tissue overlying the mucosa of the medial surface of the bilateral vocal process of the arytenoid . ConservPatient 2. A 30-year-old male with traumatic brain injury after an assault required prolonged intubation. Following extubation, the patient was found to have a granuloma expanding the posterior commissure (mmissure . Transcummissure . SurgicaEndolaryngeal injuries continue to exist despite advances in endotracheal tube design and improvements in critical care management of patients on a mechanical ventilator. Mucosal injury typically occurs at one or more of the three most sensitive mucosal areas: the medial surfaces of the vocal process, the interarytenoid area, and the posterior subglottis along the inferior aspect of the cricoid cartilage .While temporary dysphonia and mucosal injury to the glottis following tracheal intubation are quite common, glottic stenosis is less common and reportedly varies from 4 to 14%, with higher rates following prolonged intubation , 6. DuraFactors such as local infection, inflammatory disorders, chronic disease states, hypotension, gastric reflux, tube mobility, and impaired pulmonary toilet are likely to be significant in the development of postintubation complications . AdditioBogdassarian and Olson divided PGS into four categories of increasing severity . Type 1 Early edema and granulation tissue usually resolve within three weeks but the authors recommend that the airway should be monitored closely for resolution of symptoms. Nonoperative treatment recommendations vary considerably and are beyond the scope of this discussion. Development of changes in voice or dyspnea on exertion or other aerodigestive complaints a few weeks to months after intubation should prompt suspicion of restrictive posterior glottic stenosis. Immobile vocal cords may be best assessed during operative direct laryngoscopy with palpation of the arytenoid complex. Laryngeal electromyography may also be used to differentiate immobile vocal folds from paralysis or other neurologic disorders.While postintubation injury is common, long-term complications are uncommon. This report demonstrates the development of PGS from postintubation granuloma, which has long been suspected but not documented. We emphasize the importance of serial examination of any patient who develops persistent voice alteration with findings of posterior glottic granulation tissue or with immobility or reduced mobility of the arytenoids after intubation. The photodocumentation shows the progression of common postintubation mucosal injury to much less common posterior glottic stenosis.


Mechanical ventilation in newborns, children and adults leads to lung injury and has been shown to induce the formation of proteinaceous lung oedema causing epithelial disruption and resulting in changes in lung perfusion. Lung injury leads to reduced compliance, worsening of shunt fraction and an inflammatory response that results from high end-inspiratory transpulmonary pressures (Ptp) and inadequate End Expiratory Lung Volume (EELV). Lung injury results if a lung is ventilated from collapsed state with each ventilator cycle. Thus the first concept of treatment in acute hypoxic respiratory failure is: “Open collapsed lung units and keep them open without overdistending them”. The open lung is one in which there is little or no atelectasis and an optimal gas exchange. Lung recruitment strategies transiently increase Ptp to reopen the alveolar units not aerated or poorly aerated but recruitable.Lung recruitment maneuvers may restore EELV resulting in more stable alveoli, and reduce shear stress associated with the alveolar cyclic opening and closing. Lung recruitment can be performed in several ways: in delivery room with a T-piece device, in the NICU setting by using a conventional or high frequency oscillation (HFO) ventilator, and is commonly achieved by increase of end-expiratory lung volume with positive end expiratory pressure (PEEP) or end inspiratory lung volume by inspiratory holds or sustained inflation. Recruitment during both, HFO and CMV , follows similar concepts when using small tidal volume ventilation but is easier to achieve at bedside during HFO.- and/or death) were not improved.The Sustained Inflation maneuver (SI) applies a high pressure to the lung for a short period (30 sec) before returning to previous mean airway pressures. This volume recruitment strategy has been shown to be as protective as high frequency oscillatory ventilation (HFO) at similar lung volumes. Infants treated with a SI in delivery room had improved short-term respiratory outcomes , but major outcomes (BPD Bronchopulmonary dysplasiaRecent reports describe improvements in arterial oxygenation with the use of recruitment maneuvers during CMV and/or HFO. However, the impact of these strategies on patient important outcomes such as survival is still unclear.Further clinical studies are needed to evaluate the efficacy of SI, including selection of patients, setting of SI maneuver in terms of duration and peak pressure, timing of surfactant administration. Further studies are also needed to elucidate the impact of recruitment maneuvers during mechanical ventilation, including survival and BPD.


We generated a novel fully human anti-EGFR antibody showed lower toxicity and higher efficacy in the preclinical, toxicological and pharmacological studies when compared to a commercial drug Cetuximab. In this research, we aimed to understand the probably mechanism.Firstly, the mAb prefers binding to higher expression of EGFR. Its binding avidity on IgG form of the mAb to EGFR was equivalent to Cetuximab, while the Fab affinity was significantly lower. As a result, Higher EGFR expression tissues like tumor obtained the equivalent efficacy as Erbitux, while normal tissues with lower EGFR expression consequently showed lower toxicity.Secondly, the new mAb had higher concentration in tumor tissue than Erbitux. The accumulation of antibody in tumor local might induce stronger ADCC effect, and resulted in higher anti-tumor efficacy than Erbitux in a Hu-WBC NOD SCID xenograft mice model.


Localised prostate cancer, in particular, intermediate risk disease, has varied survival outcomes that cannot be predicted accurately using current clinical risk factors. External beam radiotherapy (EBRT) is one of the standard curative treatment options for localised disease and its efficacy is related to wide ranging aspects of tumour biology. Histopathological techniques including immunohistochemistry and a variety of genomic assays have been used to identify biomarkers of tumour proliferation, cell cycle checkpoints, hypoxia, DNA repair, apoptosis, and androgen synthesis, which predict response to radiotherapy. Global measures of genomic instability also show exciting capacity to predict survival outcomes following EBRT. There is also an urgent clinical need for biomarkers to predict the radiotherapy fraction sensitivity of different prostate tumours and preclinical studies point to possible candidates. Finally, the increased resolution of next generation sequencing (NGS) is likely to enable yet more precise molecular predictions of radiotherapy response and fraction sensitivity. Heterogeneity in tumour biology between prostate tumours results in a varied response to radiotherapy. At present no molecular tissue biomarkers are in routine clinical use to risk-stratify patients with localised prostate cancer. Instead, current management of localised prostate cancer is based upon established clinical risk factors including presenting PSA, clinical or radiological T (tumour) stage, and the total Gleason score. However, estimates of recurrence and survival vary considerably; for example, in the National Comprehensive Cancer Network (NCCN) intermediate risk group biochemical failure at five years following definitive local therapy varies from 2% to 70% . AlthougThe lethality of radiotherapy is centred on the creation of chromosomal lesions including DNA double strand breaks (DSB), which are particularly lethal when they cluster in close physical proximity to each other . Cells tAnother important radiobiological question at present is the radiotherapy fraction size sensitivity of prostate cancer, as measured by the alpha/beta ratio. An expanding body of evidence points to the alpha/beta ratio of prostate adenocarcinoma being as low as 1.5 , suggestRecent rapid progress in next generation sequencing techniques offers huge potential for personalisation of radiotherapy treatment, despite some of the required technological expertise being currently beyond the scope of most routine pathology laboratories. Other routinely available histopathological techniques such as immunohistochemistry (IHC) or genomic techniques such as fluorescent in situ hybridisation (FISH), comparative genomic hybridisation (CGH), and polymerase chain reaction (PCR) have identified many candidate biomarkers which with further validation could rapidly enter the clinic.Molecular biomarker development following prostatectomy has progressed at an accelerated pace compared to following radiotherapy due to limited tissue availability with the latter treatment . CriticsThis paper aims to review biomarkers predicting radiotherapy response in prostate cancer incorporating genomic signatures and individual candidates as well as biomarkers identified by longer established techniques. It does not address microRNA or biomarkers involved in the diagnosis of prostate cancer or prognostication outside of radiotherapy treatment; these were comprehensively reviewed in a recent paper in this journal .IHC enables direct evaluation of protein expression, which is advantageous as proteins are determinants of cellular function. Recent comprehensive genomic and proteomic work suggests that changes in nucleic acid do not necessarily translate to corresponding changes in protein expression . IHC is The Radiation Therapy Oncology Group (RTOG) 8610 and 9202 clinical trials of radiotherapy and varying lengths of androgen deprivation for localised prostate cancer have reported several biomarkers predicting outcome using IHC . A consiPollack et al. recently modelled the risk of distant metastases using expression of the above 5 candidates plus the apoptotic proteins Bcl-2 and Bax with competing risks hazard regression, adjusting for age, PSA, the Gleason score, T-stage, and treatment . The resThe role of hypoxia markers in prognostication following radiotherapy is more controversial. VEGF and HIF1-alpha were not included in the RTOG modelling of risk of distant metastases because an earlier RTOG study failed to demonstrate a significant association of VEGF with any survival outcomes following radiotherapy . HoweverThere are several possible reasons for conflicting data regarding hypoxic biomarkers and prediction of radiotherapy response. These include differences in the size of patient cohorts, NCCN risk group, and IHC cut points used to determine high expression of VEGF and HIF1-alpha. Furthermore, our understanding of how hypoxia impacts DNA repair is evolving rapidly. Recent studies suggest that hypoxia induces downregulation of proteins within the DNA double strand break repair pathways of homologous recombination (HR) and nonhomologous end joining (NHEJ) –49. ThisWith regard to DNA repair, error prone NHEJ operates in all phases of the cell cycle to repair DNA DSB; DNA PKcs has a key role in NHEJ by forming a synaptic complex bringing the free broken ends of DNA together with other ligating enzymes. Nuclear expression of DNA PKcs using IHC showed an independent association with biochemical recurrence after radiotherapy. However, other NHEJ proteins, also evaluated with IHC, such as Ku70, Ku80, and XRCC4 were not predictive of relapse .The TMPRSS2/ERG fusion is an important cellular rearrangement occurring in 50% of localised prostate tumours and ERG protein expression using IHC has been shown to be a robust surrogate for detecting the gene fusion . HoweverThere is thought to be direct cross talk between the EGFR cellular proliferation pathway and DNA repair. This provides a possible biological rationale for the observed correlation between high EGFR expression and poor prognosis following prostate radiotherapy . HoweverThe antiapoptotic protein Bcl-2 and proapoptotic Bax both have key roles in determining cell fate following radiotherapy. Independent prediction of survival outcomes has been demonstrated in some , 23, 43, in vitro clonogenic assays rodent cell lines with defects in NHEJ showed loss of fraction size sensitivity [ in vivo irradiated human skin showed that a 10-fold increase in the use of HR to repair radiation-induced DNA DSB towards the end of radiotherapy correlated with loss of fractionation sensitivity seen clinically [Identifying biomarkers of radiotherapy fraction size sensitivity is another area of unmet need. There is a tight association between proliferative indices and fraction size sensitivity of normal tissues . Normal sitivity , 66. In inically . On averinically . HoweverUse of a fluorogen with FISH, rather than a chromogen as in IHC, means that interpreting expression can be more straightforward than with IHC . In addiFISH has been used to demonstrate a role for biomarkers of cell proliferation such as PTEN and c-myc in predicting radiotherapy response. Loss of the tumour suppressor gene PTEN and amplification of the oncogene c-myc have both been associated with inferior outcomes following radiotherapy  Table 2Table 2. PCR array involves synthesis and amplification of complimentary DNA (cDNA) prior to expression profiling. Although amplification can introduce bias, this multiplex technique is particularly useful when tissue and hence nucleic acid quantity is limited. A reduction in mRNA (messenger RNA) of the cell-cell adhesion molecule E-cadherin has recently been associated with poor outcome after radiotherapy, but not after primary androgen deprivation therapy alone using PCR array . The autCGH differs from Reverse Transcription Polymerase Chain Reaction (RT-PCR) in that it measures DNA copy number variations rather than messenger RNA expression. It thus enables a measure of genomic instability and can calculate the percentage of genomic alteration (PGA) per tumour sample. Today CGH arrays are able to evaluate global copy number variations with as little as 100 ng of DNA . This isUsing CGH, copy number loss of several novel biomarkers with diverse functions has been proposed, as well as further validation of previously identified candidates including PTEN  Table 3Table 3. A number of RNA expression signatures have been proposed to risk-stratify in localised prostate cancer , 75. TheThe first known DNA based signature to predict recurrence after EBRT has recently been reported  and was Next generation sequencing (NGS) offers enormous potential for personalisation of treatment. It enables assessment of genomic events usually affecting more than 1 kbp such as structural copy number alterations and chromosomal rearrangements including translocations, inversions, and recombination events. In addition, the powerful resolution of NGS also enables detection of events affecting less than 1 kbp such as substitutions, insertions, and deletions . To our Fundamental aspects of cancer biology including DNA repair and hypoxia are intimately related to the efficacy of radiotherapy. It is therefore not surprising that over recent decades a number of promising tissue biomarkers have been developed using a range of molecular techniques. Whilst the majority of biomarker candidates are protein markers developed using IHC, markers of genomic instability using more quantitative techniques have shown excellent prognostic capability. Validation of these biomarkers is a priority so that the added benefit to standard clinical parameters can be clearly quantified and existing inconsistencies resolved. Development of predictive biomarkers that differentiate benefit from different local treatments and active surveillance would further enhance personalised management of early prostate cancer. Challenges include the need for standardised reproducible protocols and antibodies for IHC, together with the technical limitations of using very small biopsies for genomic techniques. However, technology continues to advance rapidly and the potential for molecular biomarkers to improve prediction of both sides of the therapeutic ratio of radiotherapy for localised prostate cancer is hugely promising.


Tool, protocol feasibility, recruitment process, electronic data capture, multi-centric clinical trials.The EHR4CR central query workbench supports pharma and CRO users in various stages of the multi-centric clinical trial lifecycle ranging from protocol feasibility, over subject identification and recruitment to clinical trial execution and adverse event reporting. It offers an intuitive graphical user interface for building, managing and executing formalized eligibility criteria queries Figure  to suppoThe EHR4CR central workbench facilitates the patient recruitment process by offering workflow-driven administration and monitoring of the recruitment process for multi-centric trials. The study manager can select clinical sites of interest and invite them to participate in the study. Study metadata including protocol definition and formalized eligibility criteria queries are exchanged and synchronized with the participating clinical sites. The overall study status and individual clinical site participation status are continuously monitored and updated. The study manager is automatically informed about relevant changes occurring at each of the engaged clinical sites such as changes in the number of potential candidate patients, the number of consenting patients, patients in screening, included or excluded patients at each site.In order to support clinical trial execution workflows, the EHR4CR central workbench will provide “Retrieve Form for Data Capture” (IHE-RFD)  capabiliProtocol feasibility functionality has been evaluated in three separate UAT rounds involving specialists from pharma and 11 pilot clinical sites. Subject identification and recruitment functionality is being evaluated. Support for clinical trial execution is still in development (November 2014).Pharma, CRO .http://www.ehr4cr.eu


Gambusia) inhabiting tidal creeks across six Bahamian islands. We found that genital shape and allometry consistently and repeatedly diverged in fragmented systems across all species and islands. Using a model selection framework, we identified three ecological consequences of fragmentation that apparently underlie observed morphological patterns: decreased predatory fish density, increased conspecific density, and reduced salinity. Our results demonstrate that human modifications to the environment can drive rapid and predictable divergence in male genitalia. Given the ubiquity of anthropogenic impacts on the environment, future research should evaluate the generality of our findings and potential consequences for reproductive isolation.The aim of this study rests on three premises: (i) humans are altering ecosystems worldwide, (ii) environmental variation often influences the strength and nature of sexual selection, and (iii) sexual selection is largely responsible for rapid and divergent evolution of male genitalia. While each of these assertions has strong empirical support, no study has yet investigated their logical conclusion that human impacts on the environment might commonly drive rapid diversification of male genital morphology. We tested whether anthropogenic habitat fragmentation has resulted in rapid changes in the size, allometry, shape, and meristics of male genitalia in three native species of livebearing fishes (genus:  Humans are altering ecosystems worldwide, but the evolutionary consequences of such impacts are poorly understood Palumbi . A wide Changes in environmental conditions  commonly alter the strength or mode of sexual selection in diverse taxa  human impacts alter the environment, (ii) ecological variation frequently influences the nature or strength of sexual selection, and (iii) sexual selection drives rapid and divergent evolution of male genital morphology. Understanding whether human impacts may ultimately drive genital diversification is critical because it carries important consequences for fitness, gene flow among populations, and the formation of new species. For instance, variation in male genital shape and size affects insemination and fertilization success in various taxa  transfer sperm internally to females both cooperatively and coercively using a nonretractable, modified anal fin called the gonopodium. Gonopodium size varies greatly across poeciliid species, ranging from 20% to 70% of the body length  and directly contacts the female during copulation  Gambusia hubbsi Breder 1934 is known to inhabit the northwestern islands of the Great Bahama Bank , (ii) Gambusia manni Hubbs 1927 appears to inhabit all other islands within the Great Bahama Bank, as well as overlap with G. hubbsi on New Providence , and (iii) a yet unnamed species, Gambusia sp., inhabits the islands of the Little Bahama Bank . Bahamian tidal creeks are shallow, tidally influenced systems typically characterized by a relatively narrow creek mouth that broadens landward to wetlands dominated by Rhyzophora mangle (red mangrove). Most of the water flux in these systems arises from tidal exchange , so salinities in unfragmented systems are typically around 35 ppt and the biotic communities comprise marine taxa , Strongylura spp. (needlefish), and Lutjanus spp.  inhabiting Bahamian blue holes where similar ecological differences over thousands of years have resulted in divergent evolution of male genital morphology. Specifically, we predict smaller gonopodia with more elongate and bony distal tips in unfragmented tidal creeks and larger gonopodia with more rounded distal tips possessing greater areas of soft tissue in fragmented tidal creeks  across six Bahamian islands  in March–April 2010 using dip nets and minnow traps. For each species, we collected multiple populations from both fragmented and unfragmented tidal creeks on two separate islands  due to concern that reduced visibility might bias the survey. We measured density of piscivorous fish using underwater visual census (UVC) methods , Nassau grouper (Epinephelus striatus Bloch 1792), jacks (Caranx spp.), and lionfish (Pterois spp.). Nearly all of the predators encountered in our surveys comprised known predators of Gambusia—barracuda, needlefish, and snappers (>99%)—with other potential predators rarely observed. We used our survey counts to calculate piscivore density (#/km2) for each site. We measured mosquitofish density using quadrat surveys, in which we counted all Gambusia individuals within 20 0.25 m2 quadrats within each tidal creek. We randomly chose quadrat locations across the microhabitats where Gambusia were observed, with all quadrats >2 m apart. For these surveys, the observer approached each predesignated area, stood still 1 m from the quadrat location, and waited 1 min prior to counting fish. We used the average of these 20 quadrat counts to calculate our estimate of Gambusia density (#/m2) for each site. This method proved highly effective due to water clarity and these fishes' general tendency to be undisturbed by our presence. We measured salinity and dissolved oxygen using a YSI 85 handheld multiparameter meter , water turbidity using an Oakton T-100 turbidimeter , and pH using a Hanna HI 98128 handheld meter . All six ecological parameters were measured multiple times over multiple years within a subset of tidal creeks to estimate repeatability—all parameters exhibited significant repeatability, confirming that our snapshot estimates provided meaningful estimates for comparison among sites  and the interaction between log10-transformed standard length and fragmentation status . Because we observed significant heterogeneity of slopes across fragmentation regimes , we could not simply interpret differences in relative gonopodium size—rather, the key source of variation involved allometry. Thus, we conducted analyses specifically designed to test for among-population variation in allometric slopes.To test for shared and unique responses of gonopodium size to tidal-creek fragmentation across the species and islands, we conducted a GLMM as described above, but with logTo directly investigate potential differences in scaling relationships among populations, we calculated allometric slopes for each population with a sample size >5 . This excluded three populations. Because it remains unclear whether slopes calculated using ordinary least squares (OLS) or reduced major axis (RMA) regression is most appropriate for allometric studies . Gonopodial distal-tip shape was quantified for 235 fish . We captured three to five images of each gonopodial distal tip at 128× magnification and stacked the photographs to comprise one composite image using Helicon Focus software  to test for shared and unique effects of habitat fragmentation on gonopodial distal-tip shape across species and islands. The 15 relative warps served as response variables. Log-transformed gonopodial distal-tip centroid size, residual gonopodium size (from log-log regression of surface area on standard length), and log-transformed standard length were included as covariates to control for multivariate allometry, as all three factors could exhibit allometric effects on gonopodial distal-tip shape. These three potential sources of allometry exhibited low multicollinearity ; VIFs <10 are not considered problematic . We used this multivariate axis to visualize gonopodial distal-tip shape differences between fragmentation regimes in tpsRelw. Individual scores along d were employed in remaining analyses involving gonopodial distal-tip shape.To obtain a single value that described gonopodial distal-tip shape for each individual, we calculated scores along n = 235). We employed GLMs for analysis of meristics because the counts  met assumptions of normality. We investigated shared and unique effects of habitat fragmentation on gonopodial distal-tip meristics across species and islands using separate GLMMs. Model terms included in each model mirrored that described above for gonopodial distal-tip shape .In addition to investigating potential divergence in multivariate gonopodial distal-tip shape, we wished to discover whether any traditional meristic characters on the distal tip have shifted as a result of habitat fragmentation. We counted the number of serrae on gonopodial fin ray 4a and the number of spines on gonopodial fin ray 3  from disGambusia density, salinity, turbidity, pH, and dissolved oxygen. For each environmental factor, our model included our five terms of interest. Consistent differences between fragmentation regimes were evident for piscivore density, Gambusia density, and salinity and were suggestive for turbidity , confirming that fragmentation status was not confounded with body size . Male Gambusia from fragmented tidal creeks exhibited a marginally nonsignificant trend toward possessing smaller gonopodia than counterparts in unfragmented sites , while the second best model included only salinity . These environmental factors tended to explain 50%  to 60% (Gambusia density) as much variance as a term for ‘fragmentation status,’ suggesting that much of the observed allometric differences between fragmentation regimes ultimately derived from some aspects of Gambusia density and salinity.Explicit examination of mean population allometric slopes revealed significant differences in allometry between fragmented and unfragmented tidal creeks using both ordinary least squares and reduced major axis regression approaches Table . Populates Table . The topd, mosquitofish exhibited more rounded gonopodial distal tips with larger areas of soft tissue in fragmented tidal creeks, while males in unfragmented sites exhibited more elongate gonopodial distal tips with more densely positioned bony segments  were largely attributed to differences in piscivore density and salinity  and salinity . The second-best model found significant effects of piscivore density alone . These environmental factors explained 65% (top model) to 67% (second model) as much variance as a term for ‘fragmentation status.’ Thus, some aspects of piscivore density and salinity appear responsible for the majority of observed differences between fragmentation regimes in gonopodial distal-tip shape.Model selection revealed that differences in gonopodial distal-tip shape between fragmentation regimes , multiple aspects of male genital morphology exhibited consistent differences across six Bahamian islands, with some changes matching our Gambusia species that found males from environments experiencing reduced predation risk evolved larger gonopodia, resulting from trade-offs between natural and sexual selection under different ecological conditions . Differences in salinity might also indirectly contribute to this pattern if correlated with other unmeasured factors associated with resource availability, such as copepod or zooplankton density. Variation in population density or salinity could affect the relative strengths of selection on gonopodium and body size, potentially mediated by female preference or locomotor requirements for foraging and social/sexual behaviors, which could also contribute to allometric differences between populations  confirms that genital shape can indeed affect sperm transfer, at least during forced copulations  than those for altering overall distal-tip shape. However, multiple sources of allometry were associated with both number of serrae and spines on the gonopodial distal tip, and serrae number differed among species. Although we have little functional understanding of serrae and spine number, these meristics have often proved useful in taxonomic work in Gambusia fishes , our study revealed that habitat fragmentation ultimately resulted in consistent differences in multiple aspects of genital morphology. While we uncovered differences among species in some traits  and among islands for some traits , the magnitude of differences between fragmentation regimes was typically as large or larger than these differences, and the nature of differences between fragmentation regimes did not significantly differ among species or islands. This suggests that human-induced ecological changes caused similar and strong selection on male genital morphology in fragmented tidal creeks and that population responses to similar selection were rapid, general, and repeatable.Gambusia affinis occurred over a roughly similar timescale as examined here (Langerhans et al. Although we did not test for a genetic basis for differentiation in gonopodial morphology in tidal-creek mosquitofish, previous work has demonstrated a genetic basis to population differences in gonopodial distal-tip shape and gonopodium size in congeners (Langerhans et al. Gambusia inhabiting fragmented and unfragmented tidal creeks have led to increased levels of reproductive isolation has not yet been investigated, but would provide valuable insight into the capacity of human impacts on the environment to promote the formation of new species on ecological timescales.Rapid differentiation of male genital morphology might result in reproductive isolation between ecologically dissimilar populations by inhibiting gene flow. Genitalia are vital for successful reproduction in internally fertilizing organisms, and mechanical and sensory incompatibilities might affect sperm transfer and fertilization success. Genital differences represent the first proposed mechanism of speciation Dufour , and to Drosophila buzzatti (Soto et al. Drosophila mediopunctata (Andrade et al. How general is the phenomenon of rapid genital differentiation associated with human-induced environmental changes? Although not yet a major focus of research, the idea that variation in ecological factors can affect the mechanisms of sexual selection influencing genital evolution is gaining traction (e.g., Rowe et al.