Pneumatosis cystoides intestinalis is a rare gastrointestinal complication in the course of connective tissue diseases, especially in scleroderma, that can lead to pneumoperitoneum or obstruction. Findings on plain radiography may reveal radiolucent linear or bubbly circular air bubbles in the bowel wall, with or without free gas accumulation in the peritoneal cavity. Treatment of pneumatosis cystoides intestinalis ranges from supportive care to laparotomy. Pneumatosis cystoides intestinalis (PCI) is a rare gastrointestinal complication in the course of connective tissue diseases (CTD), especially in scleroderma. Pneumatosis cystoides intestinalis was first described by Du Vernoi in a cadaveric dissection in 1783. This complication is characterized by the presence of multiple gas-filled cysts in the submucosa and/or subserosa wall of the intestine . AlthougHere we describe a case of pneumatosis cystoides intestinalis in the setting of scleroderma.A 65-year-old female presented with leaking fluid and pain around her jejunostomy (J) tube. She had a long history of scleroderma requiring J tube placement for supplemental feeding. On examination, there were pain and tenderness around her J tube. Initial X-rays were done to assure proper location of her J tube and demonstrated contrast contained within loops of bowel; however, the bowel wall had a “mottled” appearance, and incidental note was made of pneumoperitoneum . Follow-The patient's past medical history consisted of presenting to another hospital with acute abdominal discomfort. She had a CT scan done that showed air in the peritoneal cavity suggesting a perforated bowel. She went for an exploratory laparotomy which showed no evidence of bowel rupture. A second exploratory laparotomy was done to ensure a correct diagnosis and also found no evidence of bowel rupture. The patient has a history of a severe form of scleroderma affecting the bowel with thinning of the wall and air dissection into the bowel wall  and with resultant air leaking into the peritoneal cavity. PCI is a consequence of bowel involvement in patients with scleroderma and can cause pneumoperitoneum with bowel cyst rupturing into the peritoneal cavity, mimicking bowel perforation.Pneumatosis cystoides intestinalis (PCI) is a rare condition characterized by the presence of air-filled cysts present in the bowel wall and mesentery and may occur anywhere in the gastrointestinal tract. The cysts (0.5–10 cm in size) are found most frequently in the terminal ileum and rarely in the proximal small bowel, stomach, and colon . It is aIntestinal involvement in progressive systemic sclerosis, dermatomyositis (DM) or polymyositis (PM), and mixed CTD may be complicated by PCI. The pathogenesis of PCI is not fully understood, but several mechanisms have been suggested. Mucosal break down from steroids and other immunosuppressive agents cause Peyer's patches in the bowel wall to shrink, leading to an alteration of mucosal integrity and the potential for air dissection. These immunosuppressive agents also impair tissue repair mechanisms, further exacerbating ulceration and bowel necrosis. The bacterial theory suggests that bacterial fermentation of carbohydrates within the gastrointestinal tract leads to excessive gas formation, inducing absorption of this gas into the bowel wall . The bacGastrointestinal involvement in scleroderma is characterized by atrophy of the muscularis propria and its replacement by collagen tissue. Manometric and electrophysiological studies have shown evidence of a neuropathy of the enteric nervous system in the early stages of the disease, resulting in disturbances of digestive peristalsis and leading to gastroparesis, bacterial overgrowth of the small intestine, or constipation. Although any part of the gastrointestinal tract may be involved, the organs involved in decreasing order of frequency are the esophagus, small bowel, colon, and stomach . Suggestive findings on plain radiography comprise different pattern of radiolucent linear or bubbly circular air bubbles in the bowel wall, with or without free gas accumulation in the peritoneal cavity. CT imaging is the gold standard procedure for the diagnosis of PCI. Furthermore, CT allows the detection of additional findings that may suggest an underlying potentially worrisome cause of PCI such as bowel wall thickening, altered contrast mucosal enhancement, dilated bowel, soft tissue stranding, ascites, and the presence of portal air.Treatment of pneumatosis cystoides intestinalis ranges from supportive care to laparotomy. PCI is often benign and only conservative treatment, and followup is warranted. Surgery is generally indicated in patients with severe pain. The decision to proceed with explorative laparotomy must be based on the thorough analysis of a detailed history, physical examination, laboratory tests, and radiological studies.


How neurons process their inputs crucially determines the dynamics of biological and artificial neural networks. Synaptic input is typically considered to be merely transmitted linearly or sublinearly by the dendritic compartments. Yet, single-neuron experiments report pronounced supralinear dendritic summation of sufficiently synchronous and spatially close-by inputs. Here, we study its influence on single neuron responses and the performance of associative memory networks. First, we compute the effect of random input to a neuron incorporating nonlinear dendrites. This approach is independent of the details of the neuronal dynamics. Second, we use those results to study the impact of dendritic nonlinearities on the network dynamics in a Hopfield-type associative memory model, both numerically and analytically. We find that dendritic nonlinearities maintain network convergence and increase the robustness of memory performance against noise Figure . Interes


Drosophila melanogaster provides a powerful genetic model to gain insight into the molecular basis of nervous system development and function. The nervous system of the larvae is again a magnitude simpler than its adult counter part, allowing the genetic assessment of a number of individual genetically identifiable neurons. We review here recent progress on the genetic basis of neuronal plasticity in developing and functioning neural circuits focusing on the simple visual system of the Drosophila larva.The remarkable ability of the nervous system to modify its structure and function is mostly experience and activity modulated. The molecular basis of neuronal plasticity has been studied in higher behavioral processes, such as learning and memory formation. However, neuronal plasticity is not restricted to higher brain functions and it may provide a basic feature of adaptation of all neural circuits. The fruit fly  Drosophila melanogaster.The nervous system displays the remarkable ability to change in response to a continuously altering environment. In such a manner, internal and external stimuli remodel neuronal anatomy and function. This adaptive and dynamic process is known as neuronal plasticity. Neuronal adjustments make use of different cellular and molecular mechanisms. At the genetic level, differential gene expression can lead to changes in protein synthesis related to cytoskeletal and synaptic structure. Neuronal plasticity can be reflected by functional changes in ionic transport and electrical activity, a process known as functional plasticity. Structural modifications may result in neuroanatomical changes including circuitry remodeling. Therefore, understanding mechanisms underlying neuronal plasticity provides important insights into global brain function. Here, we review recent progress concerning neuronal plasticity in a readable nervous system, the larval brain of the fruit fly Modifications of synaptic strength and structure provide basic mechanisms underlying the formation of memories . The cycDrosophila larva is simple in comparison to its adult counterpart, containing only about one-tenth of neurons in its nervous system. However, the fly larva is able to perform complex and simple stereotypical behaviors. A conspicuous innate behavior of Drosophila larva is its aversion to light  of the larval eye . The larval eye is situated anteriorly and lateral, at the tip of the larva, associated with the cephalopharyngeal skeleton . Four PRs express exclusively rhodopsin5 (rh5) and detect blue light, the remaining eight express only rhodopsin6 (rh6) detecting green light , a small extraretinal light-sensing organ required for some aspects of circadian rhythm control . During abundant . Similarabundant , while iabundant . Thus, dFigure 1D). Increased light exposure results in a shortage of individual dendritic length and developmental delay of the third instar larval stage (L3). On the contrary, under constant darkness (DD) the length of dendritic arbors of individual LNs increases  have linked ion channels and signaling transducers with the cAMP pathway and its role in synaptic plasticity (Rac (Notch (babos-1 (de novo synthesis or degradation of cytoskeleton and synaptic proteins. Experience dependent plasticity in invertebrate models such as the fruit fly is likely to provide deep insights and novel mechanisms of how the environment models the nervous system.Neuronal plasticity is a prominent process during development, environmental experience and learning. The cAMP pathway seems to be a common feature controlling the neuroplastic process, but how the cAMP pathway is controlled in distinct contexts is still an open question. A first approach is to investigate the role of developmental hormones, ions and neurotransmitters and transmembrane proteins for the activation of the specific pathway for neuronal plasticity. Recent studies at the asticity , b. In tity (Rac , Notch (c (Notch , and bab(babos-1 . HoweverThe 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.


The historical patterns of opiate use show that sources and methods of access greatly influence who is at risk. Today, there is evidence that an enormous increase in the availability of prescription opiates is fuelling a rise in addiction nationally, drawing in new initiates to these drugs and changing the geography of opiate overdoses. Recent efforts at supply-based reductions in prescription opiates may reduce harm, but addicted individuals may switch to other opiates such as heroin. In this analysis, we test the hypothesis that changes in the rates of Prescription Opiate Overdoses (POD) are correlated with changes in the rate of heroin overdoses (HOD). ICD9 codes from the Nationwide Inpatient Sample and population data from the Census were used to estimate overall and demographic specific rates of POD and HOD hospital admissions between 1993 and 2009. Regression models were used to test for linear trends and lagged negative binomial regression models were used to model the interrelationship between POD and HOD hospital admissions. Findings show that whites, women, and middle-aged individuals had the largest increase in POD and HOD rates over the study period and that HOD rates have increased in since 2007. The lagged models show that increases in a hospitals POD predict an increase in the subsequent years HOD admissions by a factor of 1.26 (p<0.001) and that each increase in HOD admissions increase the subsequent years POD by a factor of 1.57 (p<0.001). Our hypothesis of fungibility between prescription opiates and heroin was supported by these analyses. These findings suggest that focusing on supply-based interventions may simply lead to a shift in use to heroin rather minimizing the reduction in harm. The alternative approach of using drug abuse prevention resources on treatment and demand-side reduction is likely to be more productive at reducing opiate abuse related harm. Opiates, the paradigmatic drugs of addiction, have caused medical concern for over a century and accidental self-poisoning for much longer Emblematic of this new cycle of opiate misuse is the dramatic rise in unintentional drug poisonings which currently represent the second leading cause of injury related death in the United States and the leading cause of death for 35 to 54 year olds Most of the current cases of opiate-related overdose can be traced to two fronts: that of illegal heroin consumption and the illicit use or misuse of prescription opioids, such as hydrocodone or oxycodone The few recent studies that have investigated the dynamics of opiate-related overdose have focused solely on the emergence of the prescription opioid-related overdose epidemic. Evidence from the Drug Abuse Warning Network (DAWN) data shows that there has been a significant increase in the number of Emergency Department admissions associated with oxycodone and hydrocodone Further evidence of a changing dynamic of opiate abuse comes from CDC reports suggesting that prescription opioid-related overdoses rates are increasing the most in middle age individuals OxyContin, the brand name long-acting form of oxycodone, have been linked to a self-reported rise in heroin use intertwined.There is contemporary evidence of an evolving flux between prescription opioid use and heroin use in urban areas. Ethnographic work from Montreal shows that individuals historically at risk for heroin injection have begun shifting to injecting prescription opioids While the increase in prescription opioid-related overdoses has been widely reported, the bulk of this documentation has occurred in government publications with few national longitudinal studies detailing the changing demographic characteristics of opiate-related overdose cases. Furthermore, no studies to our knowledge have investigated the relationship between prescription opioid-related overdoses and heroin-related overdoses using national data. The aim of this paper is to document changes in the demographics and examine the association between prescription opiate and heroin-related overdoses.This study uses a consecutive series of nationally representative samples of hospital admissions from 1993 to 2009 to understand the changing demographics of prescription opioid-related overdoses (POD) and heroin-related overdoses (HOD) and to measure associations between changes in POD and HOD in hospitals. Hospital-based admission data have the benefit of focusing only on severe cases that have been vetted, generally in an emergency room, using more uniform standards and better ICD-9 coding of conditions than emergency room admissions. While more uniform, hospital admissions limit the sample to only to most severe cases. The large representative national sample allows for precise estimates of the extent of hospital admissions related to opiates in both urban and rural areas and among different demographic groups.Data come from the Nationwide Inpatient Sample (NIS), Healthcare Cost and Utilization Project (HCUP), Agency for Healthcare Research and Quality SAS Software version 9.2 was used to code the data. For the analysis of trends in the demographics of POD and HOD, SAS Proc Surveyfreq was used to estimate the number of admissions adjusting the estimates for the sampling design using weights, strata and sampling units. SAS Proc Reg was used for the regression analysis of trends and SAS Proc Means was used if there was no statistically significant trend in the rates.2) and cubic (time3) coefficients estimated. A single linear time coefficient represents a straight line. Quadratic and cubic coefficients allow the slope of the line to change over time. Quadratic coefficients allow for a single rate of change in the slope of the line such as an increase in the rate of overdoses or a decrease in the rate of overdoses. A cubic coefficient allows for wave-like changes in the slope of the line, suggesting alternating periods of increasing, flat and decreasing rates. Means are only shown when the time trends were nonsignificant. When two or more of the linear, quadratic and cubic models were statistically significant the models with the highest R2 are showed. Statistical significance for the quadratic or cubic models represents the combined statistical significance of all the time variables. Means and standard deviations are reported in cases where there are no statistically significant trends.Rates of POD and HOD hospitalizations per 100,000 US population were estimated for years 1993 through 2009. Rates based on cross tabulations of POD and HOD with race, gender, and age categories in five-year increments were calculated per 100,000 members of the respective demographic group per year between 1993 and 2009. We used a weighted moving mean smoothing with the prior year and post year weighted as a one and the current year weighted twice for all the HOD data displayed in the figures. The smoothing was performed to reduce the noise in the HOD data that made it difficult to see underlying trends. We used raw HOD data for all the other analyses. These rates were used to test for linear and quadratic trends over time. A nested series of models were estimated for each series of rates. All year means were estimated, followed by a sequence of regression models with a linear, quadratic  count of HOD and POD, along with time and the urbanicity of the hospital as covariates. Urbanicity of hospitals is constructed by HCUP from the US Department of Agriculture urban rural coding The HOD model uses a one year lagged count of hospital POD, urbanicity of the hospital and interaction between urbanicity and POD, while controlling for the a one year lagged count of HOD and secular trends using a time coefficient. This model suggests that there is a strong relationship between PODs and HODs. Holding all other variables constant, each additional OD in the previous year’s hospital count of POD increases the hospital’s rate of the subsequent year HOD by a factor of 1.26 (p<0.001). A standard deviation increase in the prior year’s PODs, roughly equivalent to 7 overdoses, increases the mean expected count of HODs for a hospital by 193%. As expected urban hospitals have higher rates of HOD compared with rural hospitals by a factor of 8.42 (p<0.001) holding all other variables constant. As with the POD model, the influence of the one-year lagged PODs on the current year’s count of HODs is 15% (p<0.001) greater in rural hospitals compared with urban hospitals.The findings presented here are consistent with the existing literature describing the exponential growth of POD between 1993 and 2009. However, we also document the recent increase in HOD and provide initial evidence suggesting a link between the growth of PODs and the recent surge of HODs. While preliminary and not evidence of a causal relationship, the findings presented here provide empirical support for a growing concern about the commingling of prescription opioid and heroin abuse.Consistent with the literature, we find changes in the populations most at risk for this recent opiate-related overdose epidemic. The most dramatic increases in POD are seen among whites, women (with rates exceeding that of men) and the middle-aged. Consistent with the hypothesis that prescription opioid use is creating new users of heroin, the race-adjusted rate of HOD has doubled among whites, exceeding in 2008 that of African-Americans for the first time, while also significantly increasing among 20–24 year-olds – an age group at high risk for escalating substance abuse. The rate of HOD has declined among African-Americans, perhaps reflecting generational resilience following the devastating effect of the 1960–70′s heroin epidemic on African-American communities.The acceleration in HOD admission rates after 2006 raises fresh concerns. Previous literature declared that HOD was mostly flat during the unfolding of the prescription opioid epidemic While consistent with the above-presented explanation other hypothesis could explain these findings. For example, heroin use may follow a cyclical pattern with generational resilience and generational forgetting (i.e. the consequences) alternating. Various socio-economic causal mechanisms may also simultaneously be responsible for both forms of opiate epidemics with little need of intertwining between them. Furthermore, hospital data undercount overdoses that result in deaths or are otherwise treated in the community. These nonhospital deaths may have a different racial or ethnic composition due to differences in community risk and other behavioral factors OxyContin was approved in 1995 and sales rose rapidly, increasing 69-fold from $44.8 million in 1996 to $3.1 billion in 2010 OxyContin to physicians and responses to the prescription opioid misuse problem have been growing in number and strength since 2003 OxyContin) to which it plead guilty to falsely misrepresenting the addictive qualities of the drug compared with other pain medications However, this recent experience is consistent with the historical evidence suggesting that changes in the form of opiates, their availability and the stigma attached to them coincides with changes in the population at risk for abuse and overdose. The recent rise in POD corresponds with the rise in prescription opioid availability. This in turn has been driven by a number of factors: increasing supply, pain management advocacy, pharmaceutical marketing directed to prescribers, and drug diversion. Drug diversion is a particular problem exacerbated by doctor shopping, internet sales, theft, and improper prescribing Reversal of the “pendulum swing” from under-treatment of pain to over-treatment may have intended  and unintended consequences  OxyContin OC (crushable form) cost ∼$0.50 per mg (unpublished data) and heroin has had a mean street-price of $0.56/mg-pure OxyContin is now less desired as a street drug in its new tamper-resistant formulation Preliminary observations from a companion qualitative study in inner-city Philadelphia reveal some of the market incentives for users transitioning from prescription opioids to heroin. Heroin is inexpensive and pure by historical standards and long-acting oxycodone is typically more expensive than heroin There are several limitations to this study. The reliance on hospital coding for accurate diagnosis of the cause of opiate-related overdose induces some unaccountable variability. For example, hospitals may use a single code for all opiate-related overdoses, combining heroin and prescription opiates, making it difficult to distinguish specific drugs responsible for an overdose admission. Using hospital admissions rather than emergency room admissions also may affect the population by only selecting cases with more serious medical conditions. This sampling frame may result in a population that is older and with a higher prevalence of comorbid medical conditions than the opiate using population in general. Many opiate-related overdoses can be treated effectively in the ED regardless of the amount of opiate consumed and different hospitals may be more effective in treating ODs in the ED than other hospitals. We also cannot distinguish between overdoses resulting from illicit use, misuse or accidental use. Finally, using hospital admissions rather than individuals as the units of analysis we cannot control for individual level covariates or examine repeat visits in the lag models. While the degree to which the biases are unknown suggests caution in interpreting the results, the large representative sample of hospitals and random nature of many of these potential sources of errors suggests that the effect of these limitations is likely modest.Given the dramatic increase in the rate of POD and the strong potential for current structural reforms aimed at reducing prescription opioid misuse to inadvertently shift a proportion of prescription opioid users to heroin, a robust public health response is necessary. Active surveillance looking for a rise in heroin use/users is a good starting point. Public health measures that can address and reduce overdose deaths should include primary overdose prevention e.g. treatment of substance use, including opiate substitution therapies i.e. methadone and buprenophine. Buprenorphine, a partial opiate agonist successful in treating heroin dependency Secondary prevention efforts should include national opiate overdose awareness campaigns; these could be coupled to national policies addressing prescription drug abuse


Alzheimer's disease has become a growing socio-economical concern in developing countries where increased life expectancy is leading to large aged populations. While curing Alzheimer's disease or stopping its progression does not appear within reach in a foreseeable future, new therapies capable of delaying the pathogenesis would represent major breakthroughs.The growing number of medical benefits of cannabinoids, such as their ability to regulate age-related processes like neuroinflammation, neurogenesis and memory, raise the question of their potential role as a preventive treatment of AD.To test this hypothesis, epidemiological studies on long term, chronic cannabinoid users could enlighten us on the potential benefits of these compounds in normal and pathological ageing processes. Systematic pharmacological (and thus more mechanistic) investigations using animal models of Alzheimer's disease that have been developed would also allow a thorough investigation of the benefits of cannabinoid pharmacotherapy in the pathogenesis of Alzheimer's disease.The chronic administration of non-selective cannabinoids may delay the onset of cognitive deficits in AD patients; this will dramatically reduce the socio-economic burden of AD and improve the quality of life of the patients and their families. Alzheimer's disease (AD) is the most common neurodegenerative disease and accounts for the majority of diagnosed dementia after age 60. Currently available drugs only produce temporary relief from some cognitive symptoms without delaying, stopping or reversing the neuropathology. Due to poor efficacy of current treatments and the likely delay to implement future safe and efficacious treatments, there is an opportunity to develop preventive approaches based on currently available knowledge regarding the pathogenesis of AD. Neuroinflammation has attracted growing attention due to its slow progression and chronic nature, particularly during normal aging, as well as its involvement in various neurodegenerative diseases . NeuroinMost recent therapies have been designed and tested first in transgenic mice models and rarely take into account the ageing factor linked to AD development. As 95% of AD cases are non-familial and occur in the latter stages of life, basic knowledge on normal ageing processes and the way to delay them appear as a logical approach to tackle AD .The endocannabinoid system has recently raised a great deal of interest in AD research, notably as a powerful modulator of neuronal activity (i.e. glutamatergic neurons) or inflammatory processes -11. CannHence cannabinoids are endowed with unique proclivities that warrant their use in the prevention of age-associated cognitive decline.From the above, we postulate that modulation of the endocannabinoid system in recently diagnosed AD patients by daily administration of low-doses of cannabinoids could at minimum delay the disease progression. In the long run, a preventive approach aimed at the general ageing population may become appropriate.Epidemiological studies have already been conducted on cannabis users, in particular in adolescents for the still controversial role of cannabinoids in the development of psychosis such as schizophrenia . The pop9-THC, HU-210, anandamide, or inhibitors of endocannabinoid degradation) in mouse models of AD. Indeed, mice have been genetically engineered to incorporate human mutations of genes encoding the amyloid precursor protein (APP) and presinilins (PS) linked to familial cases of AD, and/or the hyperphosphorylation of Tau protein [Further studies should be carried out on animals to address preclinical questions. These should involve chronic administration of different non-selective cannabinoids  remain to be explored.- The expression of APP and its catabolites Aβ and sAPPα, as representatives of the amyloidogenic and non-amyloidogenic pathways, respectively. It will be of particular interest to evaluate the ratio of intracellular vs extracellular Aβ, and Aβ oligomers vs fibrils, considering the specificities of their toxicity.- Tau protein hyperphosphorylation leading to formation of neurofibrillary tangles.- Neurogenic processes that could slow or compensate ongoing neurodegeneration.- Cognitive abilities tested using a variety of behavioral tasks.The complex nature of AD advocates for the use of a multimodal drug approach that could protect from the various processes underlying neurodegeneration and thus, at minimum, delay the pathological process. The expected benefit from a chronic treatment aimed at stimulating the endocannabinoid system is a delayed progression of AD: i.e. reduced inflammation, sustained potential for neurogenesis, reduced accumulation of Aβ, reduced hyperphosphorylation of Tau and delayed memory impairment. Such results could lead to new therapeutic strategies that target both the chronic inflammation and the decline in neurogenesis associated with both normal ageing and AD. Most importantly, delaying disease progression will significantly reduce the number of severely impaired AD patients and thus reduce the growing socioeconomic burden associated with this disease.The authors declare that they have no competing interests.YM, KB, GLW, MK and SR participated to the conception of the present hypothesis. All authors drafted and approved the final manuscript.


Culex tritaeniorhynchus is the primary vector of Japanese encephalitis virus (JEV), a leading cause of encephalitis in Asia. JEV is transmitted in an enzootic cycle involving large wading birds as the reservoirs and swine as amplifying hosts. The development of a JEV vaccine reduced the number of JE cases in regions with comprehensive childhood vaccination programs, such as in Japan and the Republic of Korea. However, the lack of vaccine programs or insufficient coverage of populations in other endemic countries leaves many people susceptible to JEV. The aim of this study was to predict the distribution of Culex tritaeniorhynchus using ecological niche modeling.Cx. tritaeniorhynchus vector. Program input consisted of environmental data  and known locations of vector presence resulting from an extensive literature search and records from MosquitoMap. The statistically significant Maxent model of the estimated probability of Cx. tritaeniorhynchus presence showed that the mean temperatures of the wettest quarter had the greatest impact on the model. Further, the majority of human Japanese encephalitis (JE) cases were located in regions with higher estimated probability of Cx. tritaeniorhynchus presence.An ecological niche model was constructed using the Maxent program to map the areas with suitable environmental conditions for the Cx. tritaeniorhynchus presence provides a framework for better allocation of vector control resources, particularly in locations where JEV vaccinations are unavailable. Furthermore, this model provides estimates of vector probability that could improve vector surveillance programs and JE control efforts.Our ecological niche model of the estimated probability of  Culex tritaeniorhynchus. The primary reservoirs of the virus are wading birds, with swine serving as amplifying hosts. Despite the development of a JEV vaccine, people remain unvaccinated in endemic countries and are susceptible to JEV infection. The distribution of the JEV vector(s) provides essential information for preventive measures. This study used an ecological niche modeling program to predict the distribution of Cx. tritaeniorhynchus based on collection records and environmental maps . The model showed that the mean temperatures of the wettest quarter had the greatest impact on the model. Of the 25 countries endemic for Japanese encephalitis (JE) endemic countries, seven possessed greater than 50% land area with an estimated high probability of Cx. tritaeniorhynchus presence. Our model provides a useful tool for JEV surveillance programs that focus on vector control strategies.Japanese encephalitis virus (JEV) is transmitted predominately by the mosquito,  Flaviviridae and is endemic to Southeast and Northeast Asia, the Pacific Islands, and northern Australia  have also been implicated as important viral transmitters Cx. tritaeniorhynchus is primarily low lying flooded areas containing grasses and flooded rice paddies, but this species can also be found in urban environments in close proximity to human populations Cx. tritaeniorhynchus habitat and increasing human risk of exposure to vector populations Japanese encephalitis virus (JEV), the causative agent of Japanese encephalitis (JE), is an arbovirus that belongs to the family ustralia  [1]. TheSwine, including domestic and feral pigs, serve as amplifying hosts of JEV in endemic areas. The proximity of human populations to pig farms, sties or feral pig populations increases the risks of JEV exposure Despite the introduction of an effective vaccine to the public in the mid-1900s, JEV remains the leading cause of viral encephalitis globally Recent developments in the field of ecological niche modeling and the development of global environmental data sets have resulted in the ability to predict the distribution of vector populations that directly relate to transmission of viruses, parasites, and fungal pathogens and impact on animal and human health. Modeling to estimate the distribution of disease vectors provides useful information in disease-endemic areas, in addition to predicting how anthropogenic changes to the environment will affect disease presence Cx. tritaeniorhynchusIn the current study, the Maxent ecological niche modeling program was utilized to model the distribution of the primary vector of JEV, Cx. tritaeniorhynchus records were identified by performing a literature search in PubMed for all previous field collection studies. When exact geographical coordinates were not provided, locations were approximated by searching for the given city, town, or village using Google Earth software. Further geographical data points for the distribution of Cx. tritaeniorhynchus were obtained through MosquitoMap (http://www.mosquitomap.org/), a database of spatial data points of mosquitoes that is maintained by the Walter Reed Biosystematics Unit, Smithsonian Support Center, Silver Hill, MD. Additional Cx. tritaeniorhynchus collection data were obtained from Force Health Protection and Preventive Medicine, 65th Medical Brigade, Yongsan Army Garrison, ROK.Geographical coordinates of known In previous modeling work in the ROK www.promedmail.org) from 1994 through 2010 , Armed Forces Research Institute of Medical Sciences, and BIKEN http://www.worldclim.org/bioclim). The WorldClim organization has processed 50 years of ground-based weather measurements to produce mean monthly minimum and maximum temperatures and precipitation in a grid format at several different resolutions. The data were further processed to produce bioclimatic variables . For this project, the highest resolution data available from WorldClim (approximately 1 km) were downloaded. In addition to bioclimatic variables, global elevation data obtained from WorldClim was re-sampled to 1-km resolution from NASA's Shuttle Radar Topography Mission (SRTM). Descriptions of the bioclimatic and elevation variables used for this study are listed in Cx. tritaeniorhynchus distribution, the values of each environmental layer at each site were extracted using ArcGIS . This allowed for a comparison to the known environmental and distribution limitations for Cx. tritaeniorhynchus in the literature.One kilometer resolution climate and elevation data were obtained from WorldClim . GeoCover was created by processing Landsat Thematic Mapper images to create land cover maps for most areas of the world. Each pixel within the GeoCover-LC represents 30 by 30 meters. To convert the image to a resolution that could be used in the Maxent model, ArcGIS was used to count the number of rice pixels within each square kilometer (33 by 33 pixels). Then, a rice percentage was calculated for each square kilometer  and stored in a final output image.A map of rice growing areas was created by processing GeoCover-LC (Land Cover) data from MDA Information Systems, Inc. (http://www.cs.princeton.edu/~schapire/maxent/) was utilized to model the distribution of Cx. tritaeniorhynchus based on previously obtained geographical locations. Maxent utilizes a maximum entropy algorithm to analyze values of environmental layers, such as temperature, precipitation, and elevation, at known locations of species occurrence (collection records) to estimate the probable range of the species over a geographic region The Maxent 3.2.1 modeling program  once with all variables, 2) dropping out each variable in turn, and 3) with a single variable at a time. Variables are considered import if they produce high training gains when used alone in a model. A variable is also important if the training gain is low when the variable is removed from the model Maxent utilizes two approaches to validate the accuracy of the model. The first method randomly selects occurrence points to be withheld from the model building to use as testing points. Using multiple definitions, a set of thresholds split the continuous probability values of the model into ‘predicted presence or absence’ categories. Maxent then calculates the p-value based on the null hypothesis that testing points will be predicted as “present” no better than by a random model. The second method calculates the Area Under the Curve (AUC) of the receiver operator characteristic (ROC), a graphical depiction of the sensitivity versus one (1) minus the specificity of the model often used to validate ecological niche models Cx. tritaeniorhynchus geographical locations were utilized to construct the ecological niche model  environmental layer is the environmental variable with the highest gain when used in the model by itself. The Maxent program also calculates a percent contribution for each variable in the model. The annual precipitation variable contributed 16.2% of the information used by the model, another indication that it is an important environmental factor for estimating the distribution of rhynchus . The meaCx. tritaeniorhynchus used in the model fell within 0 and 838 meters of elevation. This is consistent with the published reports that Cx. tritaeniorhynchus is rarely collected above 1,000 meters The values of each environmental variable at each recorded location of occurrence were extracted using ArcGIS . For exaCx. tritaeniorhynchus presence  possessed <1% of their total country area with a 25% probability of Cx. tritaeniorhynchus presence.ArcGIS analysis determined the approximate percentage of each country with >25% probability of nt model . Of the Cx. tritaeniorhynchus was developed using mosquito presence records, climate, and elevation variables. Locations of human cases of JE generally fell within the higher probability areas of Cx. tritaeniorhynchus  is an important contributor to the model. Additionally, the mean precipitation of the wettest quarters (bio08) and elevation also contributed greatly to the model for distribution of rhynchus . PreviouCx. tritaeniorhynchus data for China were very limited  for vector presence  increases the likelihood that the predicted Cx. tritaeniorhynchus location does not match disease acquisition location as many people travel more than a kilometer in the course of a typical day. Alternatively, additional factors other than the presence of Cx. tritaeniorhynchus may be important when determining the risk of disease. For instance, other JEV vectors may dominant in these regions estimated with low Cx. tritaeniorhynchus presence. JE is one of many febrile illnesses that affect human populations in Asia. Difficulties arise in diagnosis of JE in patients based on symptoms alone that range from mild to very severe, with laboratory tests required for confirmation. Obtaining geographical data of where human cases were acquired is made difficult due to lack of confident diagnoses, patient travel history, and spatial data. The lack of precision of the reported case locations may also contribute to lower numbers of JE cases falling within high probability Cx. tritaeniorhynchus pixels. Identification of human JE cases in this study is extremely limited and does not represent all human cases in JE endemic regions. In many cases, only a village or city name was given for reported cases. A previous study to model the distribution of Cx. tritaeniorhynchus to predict JE in the Republic of Korea found human cases to occur in areas of high estimated probability of vector presence Human JE cases were categorized based on the estimated probability of vector presence at the reported location . InteresEcological niche modeling inherently possesses limitations in that it makes predictions based solely on environmental variables that impact larval development and adult survival. Other important factors that influence vector distributions include: vector control strategies, public health campaigns, socioeconomic status, human population densities, anthropogenic changes to land (creation of vector habitat), vector species competition, and predator influences on their potential distribution and population densities. Further, the use of WorldClim data may underestimate or ignore environmental variables that occur during a short time period or transient habitat suitable for the vector to survive. Incorporation of these variables will undoubtedly increase the validity of the model. These factors are also important to take into consideration when implementing mosquito control initiatives and vaccination campaigns.The reemergence of JEV remains possible due to multiple factors. Increases in the pig farming industry, modification and expansion of arable lands for wetland rice farming, and a fraction of the population unvaccinated/non-immune, in combination with optimal climatic conditions, contribute to the potential for periodic outbreaks of JE as the one observed in the ROK


We will review the different strategies that led to the recent development of efficient viral vectors that can be successfully used to selectively transduce astrocytes in the mammalian brain.Astrocytes are now considered as key players in brain information processing because of their newly discovered roles in synapse formation and plasticity, energy metabolism and blood flow regulation. However, our understanding of astrocyte function is still fragmented compared to other brain cell types. A better appreciation of the biology of astrocytes requires the development of tools to generate animal models in which astrocyte-specific proteins and pathways can be manipulated. In addition, it is becoming increasingly evident that astrocytes are also important players in many neurological disorders. Targeted modulation of protein expression in astrocytes would be critical for the development of new therapeutic strategies. Gene transfer is valuable to target a subpopulation of cells and explore their function in experimental models. In particular, viral-mediated gene transfer provides a rapid, highly flexible and cost-effective,  Astrocytes make up most of the cells in the brain. In addition to well-characterized roles for astrocytes in regulating brain metabolism and blood flow, there is now an increasing body of evidence that astrocytes are dynamic regulators of synaptogenesis, synaptic function and network activity. This is conceptualized in the tripartite synapse model, where pre-synaptic and post-synaptic elements of neurons are surrounded and regulated by astrocyte processes .Astrogenesis occurs relatively late in development after most neurogenesis has completed . Defectsin vivo were recently developed. Additional tools that allow for temporally controlled deletion of genes, specifically in rodent astrocytes, along with improved high resolution imaging techniques, are enabling researchers to address fundamental questions in astrocyte biology for the first time. However, these tools need to be more fully expanded and exploited to better understand astrocyte biology in vivo. The situation is complicated by the recent findings that astrocytes do not represent a homogeneous cell population across brain regions as well as within the same brain region  for the production of transgenic mice has opened new opportunities to study gene expression and functions in the brain. The resulting gene expression central nervous system (CNS) atlas program GENSAT represents a powerful resource for the scientific community (http://www.gensat.org). However, it remains difficult and time-consuming to target specific cell subpopulations through transgenesis, and differences in recombination efficiency between transgenic lines complicate the analysis. We will therefore rather focus on an alternative approach to genetically manipulate astrocytes that relies on the use of viral vectors. Indeed, the development of highly efficient viral vectors for gene transfer in the CNS is providing new systems for localized and controlled gene expression. Even if such approach requires the stereotaxic injection of the viral vectors in each animal, it significantly reduces the costs of in vivo experiments, and it can be used in combination with mouse models for conditional gene targeting, providing high flexibility and versatility to replace, modify, induce, or block expression of target genes. We will therefore review the recent development in this field that led to the emergence of effective and selective viral vectors for transducing astrocytes in vivo.Yet, a very important application of transgene expression is the visualization of a large population of astrocytes Figure 1). Among the most widely used vectors for CNS applications are the lentiviral (LVs) and adeno-associated viral vectors (AAVs) which have particularly attractive properties which include, the capacity to infect non-dividing cells, the absence of cytotoxic or immune response, long-term transgene expression and large diffusion in the brain. At least for LV, the cloning capacity is sufficient to integrate most of the genes of interest  were derived from the Ad type 5 serotype . These vIn the mid 1990s, the first AAV (from serotype 2) and LVs were reported . The AAVex vivo approach was used, with the transduction of hematopoietic CD34+ cells and re-infusion of corrected cells in the patients. An immunological improvement occurred in the two treated children aged 9–12 months in combination with a blockage of the demyelinating lesions observed by magnetic resonance imaging (MRI), 12–16 months after gene therapy (http://www.oxfordbiomedica.co.uk).Finally, the most extensively characterized LVs are derived from HIV-1, which is a subclass of retroviruses. Retroviruses are lipid-enveloped particles comprising a homodimer of linear, positive-sense, single-stranded RNA genomes of 7–11 kb. Following entry into target cells, the RNA genome is retro-transcribed into linear double-stranded DNA and integrated into the cell chromatin . To decr therapy 2012. Inin vivo. However, the first generation CNS viral vectors had a strong neurotropism in vivo .However, it is important to mention that a number of parameters could alter the tropism. These include, amongst other factors, the purity of the vector, the mode of production, the site of administration, species, the developmental stage, and normal or pathophysiological conditions. Unfortunately, data gathered in primary cultures (neurons and astrocytes) are not predictive of the trocytes  while tr-alpha -alpha . The AAVer cells . The AAVer cells . Recentler cells . The ideer cells . Finallyer cells  but the  per mm3 .Figure 2). It dramatically broadens LV tropism by facilitating transduction of various cell types in different species, it stabilizes the vector particles from shear forces during centrifugation thereby allowing vector concentration and it directs LV to an endocytic pathway, which reduces the requirements of viral accessory proteins for transduction . The most used pseudotype for LV is VSV-G which confers some interesting properties to the vector  promoter reveals that 5′-flanking regions of the gene are sufficient to direct transgene expression in astrocytes . Two fraansduced , correspansduced .Figure 2). The vector LCMV/LV-gfa2-Cre was injected into the SVZ of Rosa26 mice that express the sequence LoxP-stop-LoxP-LacZ  of non-astrocytic cells is observed. In addition, most studies rely on the use of the GFAP promoter. Large initiatives are underway to characterize the regulatory elements of the whole human genome . miRNAs are small non-coding RNA of 19–25 nucleotides that mediates post-transcriptional gene suppression  machinery to obtain tissue-specific expression . Thanks to the unique features of these new viral vectors, it has already been possible to make significant advances in two areas of research related to the development of innovative therapies and the modeling of neurological disorders.In conclusion, the use of these three different strategies  has enabled the development of efficient gene transfer systems to specifically target astrocytes , two different strategies were recently used. The first one relies on the use of the Cre/loxP system to silence the expression of the mutant protein in specific cell types by crossing different Cre-expressing transgenic mice with transgenic mice expressing the mutant protein flanked by loxP sites in all cell types. The opposite strategy consists of selectively expressing the mutant protein in specific cell types using either specific promoters such as GFAP or by crossing different Cre-expressing transgenic mice with transgenic mice expressing the mutant protein after a STOP cassette flanked by loxP sites.In order to dissect out the specific role of different cell populations These two strategies were useful in providing evidence that astrocytes play a key role in the pathogenesis of ALS , spinoceGlioblastoma multiform (GBM) is the most common primary tumor developing in the brain from astrocytes. Due to the quick proliferation and its infiltrative nature, complete ablation by surgery is almost impossible. The prognosis is very poor, with a median survival of 14.6–19.6 months and an inevitable relapse within a few months after the resection . Viral-mHowever, appropriate viral vectors for the treatment of GBM are different from those developed for the treatment of neurodegenerative diseases. For GBM therapy, the aim is to mediate destruction of proliferating cells. Glial targeting is achieved either by the injection of the vector into the tumor mass, by choosing a vector which target dividing cells or having a partial tropism for glial cells, as it is the case for Ad .The first studies used a replication-deficient mouse moloney leukemia virus (MLV) that infected dividing cells and expressed a suicide gene  on the nigro-striatal dopaminergic system . HoweverAstrocyte activation, characterized by hypertrophic somata and processes, is an early hallmark in most neurodegenerative conditions. The functional impact of this activation on the progression of these diseases is still elusive and their therapeutic potential is yet unexploited. A recent study has taken advantage of the strong astrocytic tropism of AAV2/5 expressing the astrocyte-specific promoter Gfa2 to test the potential of astrocyte-targeted therapeutics in an intact animal model of Alzheimer’s disease AD; . It was in vivo. Thanks to these tools that include targeted transgenesis and viral transduction, considerable advances were made in the understanding of astroglial biology. This first generation of astrocytic viral vectors was instrumental to start depicting their role in specific brain regions of different species. However, a better determination of the numerous functions played by astrocytes during development, in adulthood and disease will require new viral vectors that can further resolve the intimate relationship between neurons and glia in the maturing brain (in vivo assessment of efficient miRT sequences will also permit one to ameliorate the detargeting strategy. Similarly, the identification of the receptors required for the binding of the viral particles to astrocyte subpopulations will represent a major step toward the production of more efficient astrocytic viral vectors. In addition to these strategies which are already used to drive the tropism of viral vectors toward astrocytes, new viral vectors could be developed. Among these emerging viral vectors, baculoviral vectors take advantage of their natural tropism for astrocytes (The growing importance of astrocytes in crucial brain functions and also dysfunctions has led to the development of new genetic tools to label and manipulate these glial cells ng brain . One impng brain  but is jtrocytes . Their ltrocytes . Clinicatrocytes . This ilThe 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.


Wilson disease is rarely reported among African children. This report describes the second case report of a Nigerian child with Wilson disease in three decades.th International Meeting of Wilson Disease and Menkes Disease, a diagnosis of Wilson disease was made.An eight-year-old African boy presented with generalized oedema and ascites and proteinuria. Over the next three weeks he developed conjugated hyperbilirubinaemia, severe coagulopathy and prominent extrapyramidal features consisting of rigidity, tremors at rest and in action, shuffling gait, slurred speech and emotional lability. Slit-lamp examination of his eyes revealed Kayser-Fleischer rings and sunflower cataracts. His serum caeruloplasmin level was 5mg/dL. Using the scoring system proposed by the 8Wilson disease does occur in African children, although the diagnosis is rarely made. A diagnosis of Wilson disease should be entertained in the evaluation of African children presenting with liver dysfunction and/or extrapyramidal neurological features. Wilson Disease (WD) is a rare autosomal recessive disease resulting in a systemic overload of copper. The reported prevalence is 30 per million people . HoweverAn eight-year-old African boy, previously well, was referred to our unit on account of nephrotic syndrome. He had presented to our emergency department a day before with ascites, facial swelling and reduced urinary output in the preceding two weeks. After two days of hospitalization, he became deeply jaundiced with worsening of the generalized oedema. A day later, he had three brief episodes of generalized clonic seizures over a two-hour period and became increasingly drowsy. The seizures were aborted with diazepam and he remained seizure-free after a short course of phenobarbitone. A physical examination did not support a diagnosis of meningitis and other aspects of a neurological examination were normal. He had ascites and non-tender hepatomegaly; his spleen was not palpable. A review of his history did not reveal chronic or recent use of orthodox or non-orthodox medicines. In addition, there was no previous or current history of similar illness in his four siblings and close contacts. Both his parents were Africans, of the Igbo ethnic group from southeast Nigeria, but were not known to be blood relatives.In the second week of hospitalization, our patient developed tremors of his hands while at rest and when reaching for objects. He became clumsy when performing chores involving the use of his hands. Subsequently, he was observed to be stiff globally, with his trunk arched forward and fisting of the hands (left more than right). His gait was noticed to be shuffling with a tendency to fall forward when trying to walk. At the same time, his face retained a wry smile and his speech became slurred and dysarthric. He frequently complained of generalized body pain and derived some relief when his clenched fist was helped open. He was also noticed to be emotionally labile; he cried inconsolably when asking for food. He was reviewed by a paediatric neurologist who pointed out the possible presence of Kayser-Fleischer (KF) rings on both eyes. A slit-lamp examination by an ophthalmologist promptly revealed the presence of both KF rings Figure  and sunfInitial investigations showed proteinuria of grade 1+ and a spot urine protein to creatinine ratio of 1. Table th International Meeting of Wilson Disease and Menkes Disease [Using the scoring system proposed by the 8 Disease , our patHis diet was restricted to high calorie, low protein and low copper meals. Vitamins, including vitamin K, and lactulose and neomycin were commenced. Control of the generalized oedema was achieved with furosemide and spironolactone. Long-term zinc and pyridoxine therapy was started. The use of trihexyphenidyl helped achieved some control of the spasticity and rigidity, sufficient to allow resumption of basic activities such as walking and holding objects. His liver dysfunction also improved, with near resolution of the international normalized ratio (from 12 to 4).WD is a rare autosomal recessive disorder resulting in copper overload. Most cases have been reported in developed countries -6. ApartChildren with WD are usually normal at birth and may remain healthy for a variable period of time; most cases present in the second and third decade of life . Our patThe neurological features of WD are primarily due to the deposition of copper in the lenticular nuclei, although areas like the brainstem and cerebellum can be affected . In our th International Meeting of Wilson Disease and Menkes Disease to make a definite diagnosis in our patient. A score of 4 or more makes the diagnosis of WD likely. This scoring system has been validated with high sensitivity and specificity as well as high positive and negative predictive values [We used the scoring system proposed by the 8e values .The long-term treatment of symptomatic cases of WD entails the chronic use of copper chelators and zinc, while liver transplantation provides a cure ,5. The cWe have described the second known case of WD in a Nigerian child, its rarity in the African population, and highlighted the varied clinical manifestations of the disease.Written informed consent was obtained from the legal guardian of the patient for publication of this case report and accompanying image. A copy of the written consent is available for review by the Editor-in-Chief of this journal.The authors declare that they have no competing interests.CIE was the primary physician of the child and initiated the writing of the case. NB extracted the clinical data from the medical record. ARS performed the slit-lamp examination while FEAL made the provisional diagnosis of WD and was a major contributor to writing the case. All authors read and approved the final manuscript.


Tumors originating from ceruminous glands are rare lesions of the external auditory canal. The lack of specific clinical and radiological signs makes their diagnosis challenging. We report the case of an exceptionally rare benign tumor, a syringocystadenoma papilliferum (SCAP), in an atypical location in the bony segment of the external auditory canal with uncommon clinical signs. The special traits of the case included the following: the most lateral component of the tumor was macroscopically cystic and a granular myringitis with an obstructing keratin mass plug was observed behind the mass. The clinical, audiological, radiological, and histological characteristics of the neoplasm are consequently presented. Intraoperative diagnosis of the epidermal cyst was proposed. The final diagnosis of SCAP was determined only by histological analysis after the surgical excision. The educational aspects of the case are critically discussed. Syringocystadenoma papilliferum (SCAP) was first described in the dermatological literature in the beginning of the 20th century as a “naevus syringadenomatosus papilliferus” . OccasioTo date, fewer than 150 case reports of ceruminous gland tumors originating in the EAC can be found in the literature . Among tThe present case illustrates an atypical location for an exceptionally rare disease.A 61-year-old woman presented to the tertiary referral Republican Vilnius University Hospital with a 4-month history of moderate ear pain, fullness, hearing loss, tinnitus, and periodic otorrhea on the right side. She had no previous history of ear infection or discharge. Otoscopically, the bony part of the EAC was fully obstructed by a single pink ulcerated polypoid mass with a hole in the center . GranulaCeruminous glands are normally located in the outer one-third to one-half of the external auditory canal skin overlying the cartilaginous region. It is generally accepted that the bony canal is devoid of such glands. In the present report, the SCAP originated from the anterior wall of the bony part of the EAC. Ceruminous glands tumors have previously been characterized as well-differentiated localized benign neoplasms that are occasionally cystic and demonstrate the papillary proliferation of glands that are histologically similar to normal ceruminous glands . Some auOccasionally, only histological examination may discover the proper diagnosis. However, when an insufficient amount of material is obtained, the intraglandular papillary structures of an SCAP, for example, may be easily missed. The type of the biopsy and its extent in cases of EAC lesions are still controversial issues. The analysis of similar cases in general does not suggest an exact algorithm for specific diagnoses, treatment opportunities, and clinical outcomes for lesions in the EAC. Preoperative incisional biopsy to establish the diagnosis can be performed to resolve differentiation problems; nevertheless, such biopsies involve a certain level of risk because all ceruminous gland tumors, including SCAPs, are thought to be potentially malignant , 15. In High-resolution computed tomography and magnetic resonance imaging are important modalities for diagnosing and differentiating masses in the EAC. Recently, Kamakura et al. presented the MR imaging characteristics of an SCAP of the EAC: intermediate signal intensities on T1- and T2-weighted images and slight enhancement on gadolinium-enhanced T1-weighted images . Judging


The motor nerve of the bi-articular rectus femoris muscle is generally split from the femoral nerve trunk into two sub-branches just before it reaches the distal and proximal regions of the muscle. In this study, we examined whether the regional difference in muscle activities exists within the human rectus femoris muscle during maximal voluntary isometric contractions of knee extension and hip flexion. Surface electromyographic signals were recorded from the distal, middle, and proximal regions. In addition, twitch responses were evoked by stimulating the femoral nerve with supramaximal intensity. The root mean square value of electromyographic amplitude during each voluntary task was normalized to the maximal compound muscle action potential amplitude (M-wave) for each region. The electromyographic amplitudes were significantly smaller during hip flexion than during knee extension task for all regions. There was no significant difference in the normalized electromyographic amplitude during knee extension among regions within the rectus femoris muscle, whereas those were significantly smaller in the distal than in the middle and proximal regions during hip flexion task. These results indicate that the bi-articular rectus femoris muscle is differentially controlled along the longitudinal direction and that in particular the distal region of the muscle cannot be fully activated during hip flexion. Some human skeletal muscles are anatomically subdivided into neuromuscular compartments, according to their architecture and/or innervation patterns Twelve healthy male subjects  participated in this study. At the time of the study, they were either sedentary or moderately active, and none had been involved in regular strength training for at least half a year. Before participation, the subjects were fully informed of the procedures as well as the purpose of the study, and gave written informed consent. This study was approved by the local ethics committee on human research, and all procedures were conducted in accordance with the Declaration of Helsinki.Each subject participated in familiarization and experimental sessions. Forty-eight to 72 hours before the experimental session, subjects attended a familiarization session in which they were asked to perform several isometric submaximal and maximal voluntary contractions of knee extension and hip flexion. The purpose of this session was to familiarize the subjects with the maximal voluntary torque production on a dynamometer  in seated position with the hip and knee flexed at 80° and 90°, respectively. In the experimental session, the subject was seated on the dynamometer, and the torso was tightly secured to the dynamometer's seat with Velcro straps. The lever arm for measurement of knee extension torque was attached 2–3 cm above the lateral malleolus, and the pad for hip flexion measurement was positioned approximately 5 cm proximal from the upper border of the patella.The surface electromyographic (EMG) signals were picked up from the vastus medialis, vastus lateralis, and rectus femoris (RF) muscles by bipolar Ag/AgCl electrodes , with band-pass filtering between 5 Hz and 3 kHz . The electrodes were placed over at the level of approximately 70%  and 90%  of the thigh length between the greater trochanter and the lateral condyle of the femur for the vastus medialis muscle, 50% and 70% for the vastus lateralis, and 30%, 50% and 70% length for RF . At all MVC) over a 1 s period with a steady torque output. Regarding M-wave data, the peak-to-peak amplitude (AmpMwave) and RMS values (RMSMwave) during the period corresponding to the area above baseline were calculated. The RMSMVC values were then normalized to AmpMwave and RMSMwave for the respective signals.The data from the trial in which the peak torque during MVC was higher between two contractions in the assessment for each task were adopted for analysis. EMG amplitude of each signal of the vastus medialis, vastus lateralis, and RF during knee extension or hip flexion MVC was quantified as the root-mean-square value  (region×task) with repeated measures was used. When a significant interaction was observed, additional one-way ANOVAs with post hoc were performed. The significance level for all comparisons was set at P<0.05. Data are expressed as means ± SD.MVC/AMPMwave and RMSMVC/RMSMwave and no significant interaction in both vastus medialis and lateralis muscles, whereas a significant interaction was observed in RF muscle  muscle activities of RF were significantly smaller during hip flexion MVC than knee extension MVC task in all regions, and that 2) there was no significant difference in muscle activities of RF during knee extension MVC among regions whereas the muscle activity of the distal region was significantly smaller than those of the middle and proximal regions during hip flexion MVC. From the present findings, it is suggested that the distal and middle-proximal neuromuscular compartments within the RF muscle exist and that individual neuromuscular compartments are separately regulated, especially during hip flexion task. It is one of the important goals in motor control researches to clarify how the central nervous system coordinates multiple neuromuscular compartments for different joint torque production. To the best of our knowledge, this is the first study to demonstrate the inhomogeneous muscle activation within RF only for hip flexion, not for knee extension task.Before interpreting the results for EMG activities of the rectus femoris muscle, a mention should be made of the methodology used in the present study. The magnitude itself of the EMG amplitude is influenced by many electrodes' position-dependent differences in peripheral factors such as muscle membrane excitability, the number of muscle fibers within the EMG electrode recording volume, skin impedance, and the position of recording electrodes relative to the innervation zone The task- and region-dependent muscle activities within a muscle have been reported in other human muscles such as the triceps surae Another possible mechanism is the inhibition by activation of the synergistic muscle(s). By using the method of post-stimulus time histogram, Naito et al. (1996) reported an oligosynaptic inhibitory reflex pathway from the brachioradialis to the biceps brachii motoneurons Previous studies have reported region-specific adaptations in muscle size of RF in response to resistance training It is well known that muscle strain injuries occur more frequently in bi-articular muscles compared with mono-articular muscles, and that especially the RF muscle and the long head of the biceps femoris muscle have greater susceptibility for the injury among the thigh muscles Despite all the precautions taken with the placement of the recording electrodes, cross-talk might have occurred as some previous studies have indicated the effect of cross-talk from other muscles on surface EMG signal of the RF muscle In conclusion, the RF muscle is activated at comparable levels along the longitudinal direction during knee extension MVC task, whereas the muscle activities are inhomogeneous within the RF muscle during hip flexion MVC, with a smaller activity in distal region compared to the middle and proximal regions. These results suggest that the central nervous system differentially controls the bi-articular RF muscle along the longitudinal direction and cannot fully activate the distal region of the muscle during hip flexion. Thus, the present findings may help in designing rehabilitation and training program for the RF muscle dysfunction due to injuries such as muscle strain.


Among the functional neuroimaging studies on emotional face processing in schizophrenia, few have used paradigms with facial expressions of disgust. In this study, we investigated whether schizophrenia patients show less insula activation to macro-expressions  and micro-expressions  of disgust than healthy controls. Furthermore, departing from the assumption that disgust faces signal social rejection, we examined whether perceptual sensitivity to disgust is related to social alienation in patients and controls. We hypothesized that high insula responsiveness to facial disgust predicts social alienation.We used functional magnetic resonance imaging to measure insula activation in 36 schizophrenia patients and 40 healthy controls. During scanning, subjects passively viewed covert and overt presentations of disgust and neutral faces. To measure social alienation, a social loneliness scale and an agreeableness scale were administered.Schizophrenia patients exhibited reduced insula activation in response to covert facial expressions of disgust. With respect to macro-expressions of disgust, no between-group differences emerged. In patients, insula responsiveness to covert faces of disgust was positively correlated with social loneliness. Furthermore, patients' insula responsiveness to covert and overt faces of disgust was negatively correlated with agreeableness. In controls, insula responsiveness to covert expressions of disgust correlated negatively with agreeableness.Schizophrenia patients show reduced insula responsiveness to micro-expressions but not macro-expressions of disgust compared to healthy controls. In patients, low agreeableness was associated with stronger insula response to micro- and macro-expressions of disgust. Patients with a strong tendency to feel uncomfortable with social interactions appear to be characterized by a high sensitivity for facial expression signaling social rejection. Given the associations of insula responsiveness to covert disgust expression with low agreeableness in healthy individuals, insula responsiveness to expressions of disgust might be in general a neural marker of the personality trait of agreeableness. The last decades have seen a rise of studies investigating abnormalities in the processing of facial affect in schizophrenia patients. Most studies agree in that patients show significant deficits in facial affect recognition The major part of neuroimaging studies in this field has used experimental paradigms including fearful faces. This seems plausible as the emotion recognition deficit for fearful faces is particularly pronounced in schizophrenia patients On the contrary, only a few neuroimaging studies have focused on facial stimuli displaying the emotion of disgust. The processing of facial expressions of disgust has been associated with another one particular brain region, namely the insula The neglect of disgust in neuroimaging studies on emotional face processing in schizophrenia is regrettable. First of all, behavioral studies have shown that patients manifest pronounced impairments in the recognition of facial expressions of disgust Another aspect warranting a closer look on disgust resides in the social dimension of this emotion. According to Rozin et al. In the case of schizophrenia patients, the social dimension of disgust is particularly interesting. On the one hand, patients often experience stigmatization because of the disease and are thus confronted with social rejection more often than healthy people In everyday communication, negative emotions are often not expressed overtly but rather occur in the form of micro-expressions, i.e. subtle and very brief expressions of an emotion The first aim of the present study was to investigate the neural correlates of the processing of micro-expressions and macro-expressions of disgust in schizophrenia patients. To our knowledge, no neuroimaging study has ever investigated the processing of micro-expressions of disgust in schizophrenia. We used a passive viewing paradigm with stimuli designed to mimic micro- and macro-expressions of facial emotions . For the purpose of the present study, the paradigm was selectively analyzed, as we focused on disgusted face stimuli and used happy faces stimuli as a control condition. A particular strength of passive viewing paradigms consists in the minimal cognitive load as cognitive load might attenuate the processing of emotional faces A second aim of the study was to test whether the neural responsiveness to facial expressions of disgust is related to social alienation in patients and in healthy controls. As outlined above, heightened insula responsiveness to expressions of disgust might be associated with increased perceptions of social rejection and thus prompt withdrawal and alienation. We assessed the degree of social loneliness and the personality trait agreeableness as indices of social alienation in our sample. Social loneliness was chosen as a direct measure of alienation, and agreeableness was chosen as a social personality trait reflecting individual differences in trusting cooperation and the tendency to be comfortable with social interactions. It has been shown for schizophrenia patients, that the frequency of social interaction was predicted by higher levels of agreeableness The study was conducted in accordance with the Declaration of Helsinki as revised in 1989 and was approved by the Ethics Committee of the University of Münster. After a complete description of the study, written informed consent was obtained from all subjects. Of the 38 psychiatric in-patients participating in the study, none were impaired in their ability to consent. This was established by the attending clinicians and senior physicians who ensured that the patients were clinically stable and able to correctly judge the potential risks and benefits of their participation. During the study description, it was ascertained that all participants could understand the aims of the study.The original sample encompassed 38 schizophrenia patients and 42 healthy controls aged between 18 and 55 years. Participants received a financial compensation. For all participants, exclusion criteria were a history of neurological disease, severe head trauma causing a loss of consciousness, substance abuse during the last six months and the usual magnetic resonance imaging contraindications. All subjects had normal or corrected-to-normal vision. Control subjects were thoroughly investigated by trained psychologists and were free of any lifetime history of psychiatric disorders, as diagnosed by the Structured Clinical Interview for DSM-IV, Axis I disorders SCID . PatientTo assess subjects' social loneliness, we administered the multidimensional loneliness questionnaire  had to be excluded from the study. The final sample encompassed 40 healthy controls as well as 36 patients. We used a passive viewing paradigm as described in other studies from our research group In both sequences, subjects were instructed to attentively watch and memorize the facial stimuli. Subjects were presented with 33 s blocks of a facial expression category  or a no-face stimulus . The no-face stimulus was used as a general baseline condition to assess task effects. In the first sequence (masked expressions), facial prime stimuli were presented for 33 ms and followed by a 500 ms mask depicting a neutral face of the same actor . In the After scanning, subjects were administered a structured protocol exploring awareness of masked stimuli. They were questioned as to whether they had noticed anything out of the ordinary during the first sequence and whether they had perceived anything before the neutral faces. Subjects reporting subjective awareness of the masked facial stimuli completed a forced-choice questionnaire concerning the emotional qualities they perceived.Images were projected to the rear end of the scanner , Magdeburg, Germany). T2* functional data were acquired with a 3-Tesla scanner  using a single-shot echoplanar sequence with parameters selected to minimize distortion in the regions of central interest while retaining adequate S/N and T2* sensitivity. Volumes consisting of 36 axial slices were acquired  212 times in a block design.Functional imaging data were motion-corrected, using a set of six rigid body transformations determined for each image, spatially normalized to the standard Montreal Neurological Institute (MNI) template with a voxel size of 2 mm edge length, and smoothed  using Statistical Parametric Mapping . Statistical analysis was performed by modeling the different conditions  as variables within the context of the general linear model convolved with a standard hemodynamic response function. Fixed-effect analyses were performed at the individual level to generate individual contrast maps, and random effect analyses were performed at the group-level. On a single subject level, contrast images were calculated comparing each emotion's face block  with the neutral face baseline.http://restfmri.net/forum/index.php), yielding an empirically determined cluster threshold of k = 37 voxels for the bilateral amygdala and a cluster threshold of k = 125 voxels for the bilateral insula.In the scope of the present paper, we focus on the disgust versus neutral contrasts. The happy versus neutral contrasts were used as a control condition for the correlational analysis. Standard univariate group-level statistics were calculated using SPM5. The insula and the amygdala were chosen as the a priori regions of interest (ROI) for the assessment of task effects and for the between-group analysis and the insula was chosen as a priori ROI for the correlational analyses. The ROIs were defined according to Tzourio-Mazoyer et al. For the between-group analysis, we entered the disgust versus neutral contrast images into a full-factorial model to assess the main effect of group and the main effect of experimental condition (masked versus unmasked disgust) as well as a group*condition interaction effect on amygdala and insula activation. Subsequent post-hoc t-tests were conducted to clarify the direction of significant main and interaction effects.In the next step, the relationships between insula responsiveness to masked and unmasked expressions of disgust and the subjects' scores on the social loneliness and agreeableness scales were investigated. We computed simple regression models as implemented in SPM5, using the scores as regressors to the disgust versus neutral as well as the happy versus neutral contrast images. To test whether correlations would differ for patients in comparison with controls and for masked versus unmasked faces, we computed interactions between the regressors and group status/experimental condition.Subsequently, the mean contrast values of significant clusters within the ROIs were extracted for each participant and further analyzed with SPSS 20 . We constructed multiple regression models predicting insula responsiveness by the social loneliness and the agreeableness scores, controlling for age, gender, trait anxiety and depression level.As patients and controls differed significantly with respect to education and intelligence, we repeated all between-group analysis using education and intelligence as covariates. As between-group effects were not smaller using the covariates, we report only the results of the original analysis.In addition to the ROI analyses, we performed exploratory whole-brain analyses which we present as supplementary material  of the social loneliness scale was high in both the patient group (α = 0.91) and the control group (α = 0.86). The sum score of social loneliness was significantly higher for the patient group, and patients showed a significantly higher within-group variance . For the agreeableness scale, reliabilities were satisfactory . Sum scores of agreeableness were significantly lower in the patient group and variances between the patient and the control group did not differ significantly . Social loneliness and agreeableness were negatively correlated in both the patient group  and the control group .21 subjects (9 patients and 12 controls) reported awareness of the facial stimuli presented before the neutral faces in the masked expressions sequence. In the subsequent forced choice questionnaire, 8 subjects (3 patients and 5 controls) correctly noticed that they had perceived facial expressions of disgust. Patients and controls did not differ significantly with respect to the subjective awareness of masked facial stimuli  or with respect to the awareness of expressions of disgust . Subjective awareness of the facial stimuli had no significant influence on neither insula nor amygdala activation in both groups.For both the masked and the unmasked expressions sequence, the task effects on bilateral insula and amygdala activation in patients and controls were assessed using the effects-of-interest F-contrast . For both sequences, we found highly significant effects of the experimental task on bilateral insula and amygdala activation in patients and controls. The group*condition ANOVA revealed a significant main effect of condition on bilateral insula activation as subjects showed higher insula recruitment during the processing of unmasked expressions of disgust . Group status had no significant main effect on insula activation but the interaction between group and experimental condition on bilateral insula activation was significant . Subsequent post hoc t-tests showed reduced bilateral insula activation in patients during the processing of masked expressions of disgust  . Figure Choosing the amygdala as region of interest, the group*condition ANOVA revealed no significant main effects of group and experimental condition. Furthermore, the group*condition interaction effect on amygdala activation was not significant.For masked disgusted versus neutral expressions, the regression analysis conducted with SPM5 yielded a significant positive correlation between the social loneliness score and right insula activation in the patient sample  . In the For the unmasked disgust versus neutral contrast, there was no significant correlation between insula activation and the social loneliness score in both groups. For the patient group, an interaction analysis confirmed that the correlation between social loneliness and insula activation was stronger for masked expressions of disgust .As for our control condition, the correlational analysis yielded no significant results. We found no positive or negative correlations between insula responsiveness to masked or unmasked happy versus neutral faces and the social loneliness score in both groups.The agreeableness score was negatively associated with bilateral insula activation to masked expressions of disgust in the patient sample  . A multiFor the unmasked disgust versus neutral contrast, agreeableness was negatively correlated with bilateral insula activation in the patient sample . In the subsequent multiple regression analysis, agreeableness was the only significant predictor of left insula activation . In controls, agreeableness was also negatively correlated with bilateral insula response to unmasked expressions of disgust . Controlling for BDI score, trait anxiety, age and gender, the association between insula response to unmasked disgust and agreeableness was no more significant (p>0.1). Instead, a strong association between neural response and BDI score emerged in controls . An interaction analysis showed that the correlation between insula response to unmasked disgust and agreeableness was stronger for controls than for patients .In patients, the negative correlation between insula response and agreeableness was stronger for masked than for unmasked expressions of disgust . In controls, there was no significant interaction between the agreeableness score and experimental condition.Using the masked and unmasked happy versus neutral contrast images in the correlational analyses yielded no significant positive or negative association between insula responsiveness and agreeableness in the patient group. In controls, we found a negative correlation between right insula response to masked happy faces and agreeableness . However, this correlation was no more significant after controlling for BDI score, trait anxiety, age and gender (p = 0.87).Disgust has been dubbed “the forgotten emotion of psychiatry” For the amygdala, no between-group difference emerged for the processing of either micro-expressions or macro-expressions of disgust. This is not surprising, as the amygdala seems not primarily involved in the processing of disgust With respect to the questionnaire data, patients had significantly higher scores on the social loneliness scale and lower scores on the agreeableness scale than controls. This parallels earlier findings of increased loneliness It should be taken into account that schizophrenia patients often face stigmatization and social rejection due to the disease Importantly, we also found a negative association between the personality trait of agreeableness and insula responsiveness to masked and unmasked disgust in healthy controls. Whereas patients showed a stronger association between agreeableness and insula activation during the processing of masked disgust faces, the opposite result emerged during the processing of unmasked disgust faces. Here, the negative correlation between insula responsiveness and agreeableness was even stronger in controls than in patients. This finding requires careful interpretation, as the correlation was confounded by depressivity in controls. However, in view of our correlational findings for schizophrenia patients and healthy controls it appears that insula responsiveness to micro-expressions of disgust might be in general a neural marker of agreeableness. Future research should follow up on this hypothesis.In our control group, we found no significant correlation between insula responsiveness to expressions of disgust and social loneliness. However, this lack of a finding might be due to reduced within-group variance in controls, as loneliness is more pronounced in clinical populations. It might be promising to investigate whether the positive correlation between insula responsiveness to disgust and social loneliness is characteristic for schizophrenia patients or also emerges in other patient groups. Future research could also test the relationship between insula responsiveness to disgust faces and other non-clinical measures of social integration in healthy controls. In particular, subjective and objective measures of experienced discrimination and social rejection might offer valuable insights. Recent studies have shown that the insula responsiveness to social rejection in a social exclusion paradigm is related to interindividual differences in discrimination-related distress The present study showed that decreased insula responsiveness to disgust in patients is related to lower degrees of social alienation. It would be important to interpret these findings in light of a general theory of insula dysfunction in schizophrenia. A growing body of research suggests a crucial role of the insula in the neuropathology of the disease. There is consistent evidence that insula grey matter volume is substantially decreased in schizophrenia patients Future research should investigate the role of insula-limbic connectivity in schizophrenia. In healthy subjects, the insula shows extensive connections with the amygdala and the anterior cingulate cortex and controls. Nevertheless, future studies are clearly needed to replicate our between-group and correlation findings before strong conclusions can be drawn. In particular, it would be important to investigate whether the same results are obtained using an event-related design. Another limitation could be seen in the high neuroleptic medication of the patient group. Replicating the study with unmedicated patients would be necessary to assure that our finding of insular hypoactivation to masked disgust in the patient group is not an effect of neuroleptic medication. Furthermore, it should be acknowledged that our correlational analyses do not justify conclusions about the direction of the relationship between social alienation and the neural responsiveness to expressions of disgust. Long-term studies could provide new evidence on how the neural responsiveness to disgust develops in the course of the disease.To our knowledge, this is the first neuroimaging study investigating the processing of micro-expressions of disgust in schizophrenia and one of few studies dedicated to the social dimension of this emotion. Despite these strengths, some limitations of the study should be noted. First of all, using a passive viewing task we had no control as to whether all subjects equally attended to the presented stimuli. This is important as schizophrenia patients often suffer from amotivation. Future research might test experimental paradigms which are more engaging but equally simple with respect to cognitive demand. Another important limitation of the study consists in the restricted ecological validity of our paradigm. We used a masking procedure to mimic rapid changes in facial expressions. Anyhow, this approach represents only a crude simulation of facial micro-expressions as mimicry in everyday communication occurs in a more fluid and less distinct way. Moreover, the backward masking procedure raises questions about the role of early visual processing deficits in schizophrenia. Schizophrenia patients show impairments in visual backward masking File S1Supporting information tables S1 – S6. Results of supplementary whole-brain analyses.(DOC)Click here for additional data file.