Can sPD-1 and sPD-L1 Plasma Concentrations Predict Treatment Response among Patients with Extraparenchymal Neurocysticercosis?

Extraparenchymal neurocysticercosis (EP-NC) is a chronic, potentially life-threatening disease that responds poorly to initial anthelmintic drug therapy. A depressed specific reactivity of peripheral lymphocytes and an increased level of specific Tregs accompanies EP-NC. The immune checkpoint pathway PD-1 and its ligand PD-L1 downregulates effector T cells, causing specific immune suppression in chronic diseases. This study explored whether their soluble forms, sPD-1/sPD-L1, are present in plasma among patients with EP-NC and if their levels could be associated with treatment response. A total of 21 patients with vesicular EP-NC and 22 healthy controls were included. Patients received standard treatment and were followed for six months to assess treatment response by assessing changes in cyst volume determined with 3D MRI. The presence of both sPD-1 and sPD-L1 was more frequently detected among patients with EP-NC than in healthy controls and had higher concentrations. Among patients, higher pre-treatment levels of both markers were associated with a poor treatment response, and the sensitivity and specificity of the sPD-1/sPD-L1 ratio for predicting any response to treatment were high. Our results are consistent with the presence of lymphocyte exhaustion and open new research perspectives to improve the prognosis of patients with this severe disease.

Low responsiveness of peripheral lymphocytes in extraparenchymal neurocysticercosis.

BACKGROUND: The morbidity and mortality of extraparenchymal neurocysticercosis (EP-NC) remain high and effectiveness of current medical treatment is suboptimal. Various factors have been implicated in the severity of EP-NC and in the poor response to treatment, but the possible role of host immune and endocrine systems has not yet been examined thoroughly. METHODOLOGY/PRINCIPAL FINDINGS: 42 participants with EP-NC before receiving standard treatment and 25 healthy controls were included in the study. Treatment response was assessed by comparing pre/post treatment parasite volumes from 3D MRI. Prior to treatment among participants with EP-NC, specific stimulation induced an increased specific proliferative response accompanied by a significant increase in IL-4, NK, NKT, Bregs and Tregs cells, whereas in healthy controls, specific stimulation induced a significant increase in IL-1ß, IL-5, CCL5, IL-6, TNF-α, NK and Bregs cells. Significant differences between participants with EP-NC and healthy controls in the specific inflammatory response were observed. Participants with EP-NC prior to treatment had significantly weaker responses of proinflammatory cytokines (IL-6, TNF-α) and NK cells, and stronger IL-4 response. Anthelmintic treatment did not promote significant peripheral immunological changes at any time, although inflammation was sustained in the cerebrospinal fluid. Serum estradiol concentration significantly decreased after anthelmintic treatment among males, and cortisol correlated negatively with IL-6 and positively with IFN-γ levels. No pre-treatment immunologic or endocrinologic parameters were significantly associated with response to treatment. CONCLUSION/SIGNIFICANCE: Prior to anthelmintic treatment, EP-NC was characterized by low lymphocyte reactivity accompanied by a regulatory response, which may be involved in the lack of peripheral immunological changes during and after treatment, although a central inflammatory response was present. This weak specific peripheral response could favor the chronicity of the infection and the poor response to treatment. Our findings highlight the need for new anti-inflammatory treatment focused on the central nervous system with less systemic immunosuppressive effects.

Taenia solium taeniasis/cysticercosis in Guatemala: a prevalent public health problem?

In Guatemala, neurocysticercosis (NCC) was first recognized in 1940; since then, cases of NCC have been reported in all Guatemalan departments. However, epidemiological studies on Taenia solium infections are scarce and most information remains unpublished. This study aims to provide evidence of T. solium infections as a public health problem in Guatemala. All information available, either published or unpublished, on T. solium infections in the country was compiled. Official data from the Ministry of Health for the period 2003-2019 were reviewed and analyzed, and all cases of T. solium infections were classified and counted. In total, 5246 cases of taeniasis and 454 cases of human cysticercosis were recorded. On the other hand, 44 studies were identified, mostly from local journals, which included 1951 cases of taeniasis, 2873 cases of human cysticercosis of which 543 were classified with complete diagnosis, and 2590 cases of porcine cysticercosis. Cases were classified by geographic region, patient sex, and Taenia species in taeniasis cases when information was available, and the departments with the highest number of taeniasis and cysticercosis cases were identified. Meanwhile, in Zacapa, a northeastern department of Guatemala with one the highest number of taeniasis cases, a young man diagnosed with a severe form of NCC and two cases of porcine cysticercosis (both confirmed by necropsy) were identified. Taken together, the data herein reported indicate that T. solium infections are a major health problem in Guatemala that needs to be addressed.

MDGAs are fast-diffusing molecules that delay excitatory synapse development by altering neuroligin behavior.

MDGA molecules can bind neuroligins and interfere with trans-synaptic interactions to neurexins, thereby impairing synapse development. However, the subcellular localization and dynamics of MDGAs, or their specific action mode in neurons remain unclear. Here, surface immunostaining of endogenous MDGAs and single molecule tracking of recombinant MDGAs in dissociated hippocampal neurons reveal that MDGAs are homogeneously distributed and exhibit fast membrane diffusion, with a small reduction in mobility across neuronal maturation. Knocking-down/out MDGAs using shRNAs and CRISPR/Cas9 strategies increases the density of excitatory synapses, the membrane confinement of neuroligin-1, and the phosphotyrosine level of neuroligins associated with excitatory post-synaptic differentiation. Finally, MDGA silencing reduces the mobility of AMPA receptors, increases the frequency of miniature EPSCs (but not IPSCs), and selectively enhances evoked AMPA-receptor-mediated EPSCs in CA1 pyramidal neurons. Overall, our results support a mechanism by which interactions between MDGAs and neuroligin-1 delays the assembly of functional excitatory synapses containing AMPA receptors.

Temporal lobe epilepsy: Evaluation of central and systemic immune-inflammatory features associated with drug resistance.

Neuroinflammation is probably one of the factors involved in drug resistance in people with epilepsy. Finding peripheral markers reflecting the intensity of neuroinflammation could be of great help to decide for which patients anti-inflammatory treatment might be an option. In this context, peripheral cytokines levels and lymphocyte phenotypes were assessed by ELISA and flow cytometry in 3 groups of subjects: drug resistant patients with temporal lobe epilepsy (DR-TLE), non DR-TLE patients and healthy controls. The same parameters were assessed in brain tissue in the DR-TLE group. Differences in the peripheral immune-inflammatory status between the 3 groups of subjects, and correlations between the central and peripheral immune-inflammatory status in DR-TLE patients were evaluated. Forty-one patients with DR-TLE, ten with non-DR-TLE and twenty controls were included. In the periphery, decrease in regulatory cells were observed in DR-TLE patients compared to controls. In addition, significant increase of IL-6 and IL-5 was observed in patients with epilepsy (particularly DR-TLE patients). Two groups of DR-TLE patients with significant differences in several central inflammatory parameters were identified in a cluster analysis. The inflammatory cluster was associated with a peripheral increase of CD4+CD38+ cells and different significant correlations between central and systemic inflammatory parameters were observed. Although their interpretation is not immediate, they demonstrate a clear dialogue between central and peripheral inflammatory reactions. In conclusion, our results add new elements to better understand the interactions between the central and peripheral compartments in patients with DR-TLE, and to help better define treatment options in this group of patients.

Cardiac Autonomic Response to Active Standing in Calcific Aortic Valve Stenosis.

Aortic stenosis is a progressive heart valve disorder characterized by calcification of the leaflets. Heart rate variability (HRV) analysis has been proposed for assessing the heart response to autonomic activity, which is documented to be altered in different cardiac diseases. The objective of the study was to evaluate changes of HRV in patients with aortic stenosis by an active standing challenge. Twenty-two volunteers without alterations in the aortic valve (NAV) and twenty-five patients diagnosed with moderate and severe calcific aortic valve stenosis (AVS) participated in this cross-sectional study. Ten minute electrocardiograms were performed in a supine position and in active standing positions afterwards, to obtain temporal, spectral, and scaling HRV indices: mean value of all NN intervals (meanNN), low-frequency (LF) and high-frequency (HF) bands spectral power, and the short-term scaling indices (α1 and αsign1). The AVS group showed higher values of LF, LF/HF and αsign1 compared with the NAV group at supine position. These patients also expressed smaller changes in meanNN, LF, HF, LF/HF, α1, and αsign1 between positions. In conclusion, we confirmed from short-term recordings that patients with moderate and severe calcific AVS have a decreased cardiac parasympathetic supine response and that the dynamic of heart rate fluctuations is modified compared to NAV subjects, but we also evidenced that they manifest reduced autonomic adjustments caused by the active standing challenge.

Taxonomic and functional responses of macroinvertebrates to riparian forest conversion in tropical streams.

Land use change threatens the ecological integrity of tropical rivers and streams; however, few studies have simultaneously analyzed the taxonomic and functional responses of tropical macroinvertebrates to riparian forest conversion. Here, we used community structure, functional diversity, and stable isotope analyses to assess the impacts of riparian deforestation on macroinvertebrate communities of streams in southern Mexico. Monthly sampling during the dry season was conducted in streams with riparian forest (forest streams), and in streams with pasture dominating the riparian vegetation (pasture streams). Samples were collected for water quality (physical-chemical variables, nutrient concentrations, and total suspended solids), organic matter (leaf litter abundance and algal biomass), and macroinvertebrate abundance and diversity. Higher temperature, conductivity, suspended solids, and chlorophyll a were detected in pasture streams, while nitrate concentrations and leaf litter biomass were greater in forest streams. Macroinvertebrate density was higher in pasture sites, while no differences in taxonomic diversity and richness were found between land uses. Functional evenness was greater in forest streams, while richness and divergence were similar between land uses, despite differences in taxonomic composition. Environmental variables were associated with taxa distribution but not with functional traits, suggesting current conditions still promote redundancy in ecological function. Isotopic analyses indicated consumers in pasture streams were enriched in 13C and 15N relative to forest streams, potentially reflecting the higher algal biomass documented in pasture systems. Isotopic niches were broader and more overlapped in pasture streams, indicating more generalist feeding habits. No significant losses of taxonomic or functional diversity were detected in pasture streams. However, changes in trophic ecology suggest landscape-level processes are altering macroinvertebrate feeding habits in streams. The changes we observed in habitat, water quality, and macroinvertebrate community were related to the removal of the riparian vegetation, suggesting the structure and function of the focal systems would benefit from riparian restoration.

Promoção da alimentação saudável no ambiente escolar: avaliação do Programa Horta Educativa em escolas estaduais de São Paulo / Promotion of healthy eating in the school environment: evaluation of the School Garden Program in state public schools in São Paulo

Introdução- O Programa Horta Educativa (PHE), coordenado pelo Fundo Social e de Solidariedade do Estado de São Paulo em parceria com a Secretaria de Agricultura e Abastecimento de São Paulo, prevê a implantação de hortas em escolas e centros infantis públicos. A horta escolar tem se mostrado uma excelente estratégia para a educação alimentar e nutricional que possibilita uma abordagem pedagógica participativa, articulando saúde, alimentação e meio-ambiente. Objetivo- Avaliar o PHE como dispositivo para a promoção da alimentação adequada e saudável (PAAS) no ambiente escolar. Metodologia- Trata-se de um estudo qualitativo de avaliação por triangulação de métodos, que avaliou o PHE em escolas estaduais do município de São Paulo. O levantamento documental e estrutural das escolas foi realizado em 18 escolas selecionadas, que desenvolveram o programa por, no mínimo, três anos consecutivos. Para a pesquisa de campo foram selecionadas duas escolas, que apresentavam a horta em plena atividade e maior envolvimento da comunidade escolar. O estudo, realizado com o emprego de diferentes métodos de investigação, foi desenvolvido na seguinte ordem: análise documental, contextualização das escolas, observação de campo, entrevistas (com professores, coordenadores e diretores) e grupos focais voltados para os estudantes. Os dados qualitativos foram analisados por meio da técnica de análise de conteúdo temática. A avaliação do PHE pautou-se pela análise do modelo lógico. Resultados- Os dados empíricos foram divididos em 4 categorias centrais: (1) O contexto alimentar escolar, abordando as especificidades de cada escola; (2) O desenvolvimento do PHE, trazendo as particularidades do projeto em ambas as escolas; (3) O ambiente escolar discutindo o papel da escola como agente de PAAS, os facilitadores e as barreiras para a PAAS na escola e; (4) A avaliação do PHE, trazendo as potencialidades, os entraves para o desenvolvimento do PHE e os resultados percebidos. Destacando as categorias de avaliação do PHE, as principais potencialidades para o desenvolvimento do PHE foram: o fato de a horta ser considerada uma atividade prazerosa para os estudantes; responsáveis engajados na realização do PHE; as parcerias estabelecidas com a escola; o envolvimento dos pais na horta e o vínculo entre o PHE e o projeto alimentação saudável das escolas. As principais barreiras ao desenvolvimento do PHE foram: escassez de recursos humanos e financeiros; não adequação do o material didático às diferentes turmas de alunos; não envolvimento dos professores e a indisponibilidade de tempo para a formação de professores. Entre os resultados percebidos, destacaram-se: mudança dos hábitos alimentares e o aumento do conhecimento dos estudantes sobre alimentação saudável; aumento no consumo da merenda escolar; valorização dos meios de produção orgânicos e promoção da educação ambiental entre os alunos; envolvimento de outras disciplinas possibilitando o trabalho interdisciplinar e maior envolvimento entre família e escola. Conclusão - O PHE, apesar das suas barreiras, promoveu a valorização e o maior consumo da merenda escolar pelos estudantes, contribuiu para a conscientização e para o aprendizado reflexivo sobre as práticas alimentares, gerando mudanças na alimentação. Políticas que envolvam a prática de hortas escolares devem ser incentivadas.

Introduction- The School Garden Program (SGP), coordinated by the Social and Solidarity Fund of the State of São Paulo in partnership with the Secretariat of Agriculture and Supply of São Paulo, provides for the implementation of gardens in public schools and public children's centers. The school garden has proved to be an excellent strategy for food and nutrition education that enables a participatory pedagogical approach, articulating health, food and the environment. Objective- To evaluate SGP as a device for the promotion of adequate and healthy food (PAHF) in the school environment. Methodology- This is a qualitative study of evaluation by triangulation of methods, which evaluated SGP in state schools in the city of São Paulo. The documentary and structural survey of the schools and the SGP was carried out in 18 selected schools, which developed the program for at least three consecutive years. For the field research, two schools were selected, which presented the active garden and greater involvement of the school community. The study, carried out using different investigation methods, was developed in 5 stages: document analysis, contextualization of schools, field observation, interviews (with teachers, coordinators and principals) and focus groups conducted with students. Qualitative data were analyzed using the thematic content analysis technique. The evaluation of the SGP was based on the analysis of the logical model. Results - The empirical data were divided into 4 central categories: (1) The school food context, addressing the specifics of each school; (2) The development of the SGP, bringing the particularities of the project to both schools; (3) The school environment, discussing the role of the school as an agent of PAHF, the facilitators and barriers to PAHF in the school and; (4) The evaluation of the SGP, bringing the potential, the obstacles to the development of the SGP and the perceived results. Highlighting the SGP assessment categories, the main potentialities for the development of the SGP were: the school garden be considered a pleasurable activity for students, tutors engaged in the implementation of the SGP, the partnerships established with the school, the parents' involvement in the garden and the articulation of the SGP with the healthy eating project in schools. The main barriers to the development of the SGP were: shortage of human and financial resources; inadequacy of didactic material to different classes of students, non-involvement of teachers and unavailability of time for teacher training. Among the perceived results, the following stand out: changing eating habits and increasing students' knowledge about healthy eating; increased consumption of school meals; valorization of organic means of production and promotion of environmental education among students; involvement of other disciplines enabling interdisciplinary work and greater involvement between family and school. Conclusion - SGP, despite its barriers, promoted the appreciation and increased consumption of school meals by students, contributed to awareness and reflective learning about eating practices, generating changes in food. Policies that involve the practice of school gardens should be encouraged.

ISSR markers to explore entomopathogenic fungi genetic diversity: Implications for biological control of tobacco pests.

Tobacco is one of the major industrial crops cultivated worldwide. Chemical control is the main method employed to reduce damage by insect pests. The use of entomopathogenic fungi represents an alternative to replace insecticides. The search for effective strains in the field constitutes a first step when developing a formulation. The objective of this work was to study genetic differences among isolates of entomopathogenic fungi obtained from tobacco grown soils using ISSR markers. The pathogenicity of the strains towards Helicoverpa gelotopoeon and Diabrotica speciosa was also assessed in order to search for a relationship between virulence and genetic diversity. Nineteen isolates were identified according to morphological features and molecular techniques as Beauveria bassiana (11) and Purpureocillium lilacinum (8). The diversity tree generated by ISSR analysis showed a high diversity among the strains. The pathogenicity towards H. gelotopoeon and D. speciosa was assessed and the logistic models generated showed that B. bassiana isolates LPSc1215 and LPSc1364 were the most pathogenic against both insect pests tested. In the diversity tree, these strains were grouped in a same cluster with a similarity level of approximately 85%, indicating a possible relationship between virulence and the band pattern generated.

Endomycobiome associated with females of the planthopper Delphacodes kuscheli (Hemiptera: Delphacidae): A metabarcoding approach.

A metabarcoding approach was performed aimed at identifying fungi associated with Delphacodes kuscheli (Hemiptera: Delphacidae), the main vector of "Mal de Río Cuarto" disease in Argentina. A total of 91 fungal genera were found, and among them, 24 were previously identified for Delphacidae. The detection of fungi that are frequently associated with the phylloplane or are endophytes, as well as their presence in digestive tracts of other insects, suggest that feeding might be an important mechanism of their horizontal transfer in planthoppers. This study draws the baseline for future research regarding mutualistic associations present in D. kuscheli as well as their physiological role in the life cycle of this important pest that might lead to developing new management strategies to keep insects populations under control.

Alternative splicing at neuroligin site A regulates glycan interaction and synaptogenic activity.

Post-transcriptional mechanisms regulating cell surface synaptic organizing complexes that control the properties of connections in brain circuits are poorly understood. Alternative splicing regulates the prototypical synaptic organizing complex, neuroligin-neurexin. In contrast to the well-studied neuroligin splice site B, little is known about splice site A. We discovered that inclusion of the positively charged A1 insert in mouse neuroligin-1 increases its binding to heparan sulphate, a modification on neurexin. The A1 insert increases neurexin recruitment, presynaptic differentiation, and synaptic transmission mediated by neuroligin-1. We propose that the A1 insert could be a target for alleviating the consequences of deleterious NLGN1/3 mutations, supported by assays with the autism-linked neuroligin-1-P89L mutant. An enrichment of neuroligin-1 A1 in GABAergic neuron types suggests a role in synchrony of cortical circuits. Altogether, these data reveal an unusual mode by which neuroligin splicing controls synapse development through protein-glycan interaction and identify it as a potential therapeutic target.

Diagnostic value of glycoprotein band patterns of three serologic enzyme-linked immunoelectrotransfer blot assays for neurocysticercosis.

The enzyme-linked immunoelectrotransfer blot (EITB) assay to detect antibodies in serum is a complementary tool for the diagnosis of neurocysticercosis (NCC). Presence of at least one glycoprotein band corresponding to a Taenia solium (T. solium) antigen indicates a positive result; however, EITB assays have multiple glycoprotein bands, and previous work has suggested that band patterns may have additional diagnostic value. We included 58 participants with a definitive diagnosis of NCC who received care at the Instituto Nacional de Neurología y Neurocirugía in Mexico City. Three different EITB tests were applied to participants' serum samples (LDBio, France; US Centers for Disease Control and Prevention [CDC]; and Instituto de Diagnóstico y Referencia Epidemiológicos [InDRE]). There was substantial variability in specific glycoprotein band patterns among the three assays. However, in age- and sex-adjusted logistic regression models, the number of glycoprotein bands was positively associated with the presence of vesicular extraparenchymal cysts (InDRE adjusted odds ratio [aOR] 1.60 p < 0.001; CDC aOR 6.31 p < 0.001; LDBio aOR 2.45 p < 0.001) and negatively associated with the presence of calcified parenchymal cysts (InDRE aOR 0.63 p < 0.001; CDC aOR 0.25 p < 0.001; LDBio aOR 0.44 p < 0.001). In a sensitivity analysis also adjusting for cyst count, results were similar. In all three EITB serum antibody tests, the number of glycoprotein bands consistently predicted cyst stage and location, although magnitude of effect differed.

Effect of dexamethasone on albendazole cysticidal activity in experimental cysticercosis by Taenia crassiceps in BALB/c mice: In vitro and in vivo evaluation.

Taenia solium is a parasite whose larvae (cysticerci) can locate in the central nervous system of humans and cause neurocysticercosis (NC). The introduction of cysticidal drugs such as albendazole (ABZ) for the treatment of NC has significantly improved its prognosis. However, treatment is not always effective, and the high levels of corticosteroids used to prevent inflammatory complications in this disease could be, partly, the cause of this observation. In this context, this study investigated, using the experimental mouse model of intraperitoneal infection with Taenia crassiceps, the influence of corticosteroid administration on the therapeutic efficacy of ABZ. We evaluated and compared the effects of ABZ, dexamethasone (DXM) and their combination (ABZ + DXM) on cyst viability, both in vitro and in vivo. Serum levels of IL-4, IFN-gamma, IL-6 and IL-10 were evaluated in the in vivo study. Results showed that the treatment with ABZ, in vitro and in vivo, was associated with a high number of parasites deaths. Concomitant treatment with DXM did not alter ABZ in vitro cysticidal activity but reduced its effectiveness significantly in the in vivo experimental model. Cytokine serum levels did not change significantly in treated mice compared to the controls. The results of this study are relevant as they indicate a negative effect of corticosteroids on the efficacy of cysticidal therapy. In human neurocysticercosis, control of inflammation is of great importance to most patients in order to avoid complications. Corticosteroids are generally used for this purpose and the results of this study demonstrate the need to find other therapeutic strategies. Further studies are needed to better understand the mechanisms involved.

Immunodiagnosis of human neurocysticercosis: comparative performance of serum diagnostic tests in Mexico.

Immunodiagnosis has a supportive role in the diagnosis of neurocysticercosis (NCC). The aim of this study was to compare the validity of seven immunodiagnostic tests among serum samples from 58 patients with NCC, 26 patients with neurological diseases other than NCC, and 15 healthy controls. One test for viable parasite detection (HP10 antigen assay) and six for antibody detection were evaluated. For the entire sample, sensitivities ranged from 55.2% (NOVALISA) to 81.0% (enzyme-linked immunosorbent assay [ELISA] Taenia solium antibody), with the sensitivity of the latter test significantly higher than that of the in-house ELISA Taenia crassiceps, NOVALISA, enzyme-linked immunoelectrotransfer blot (EITB) CDC, and HP10. Overall, specificities were high, ranging from 85.4% (ELISA Ts) to 97.1% (NOVALISA), with no statistically significant differences. Detection of HP10 antigen was significantly associated with the presence of vesicular parasites. The simple and low-cost ELISA Taenia solium antibody Ab instead of EITB is recommended to support NCC diagnosis in both rural and hospital settings in Mexico.

Factors Associated With Cysticidal Treatment Response in Extraparenchymal Neurocysticercosis.

Extraparenchymal neurocysticercosis is the most severe form of cysticercosis, and response to treatment is suboptimal. We sought to determine how demographic and clinical characteristics and albendazole sulfoxide concentrations were related to cysticidal treatment response. We conducted a longitudinal study of 31 participants with extraparenchymal vesicular parasites who received the same treatment, albendazole 30 mg/kg/day for 10 days with dexamethasone 0.4 mg/kg/day for 13 days, followed by a prednisone taper. Response to treatment was determined by parasite volumes before and 6 months after treatment. Eight participants (25.8%) had a complete treatment response, 16 (51.6%) had a treatment response > 50% but < 100%, and 7 (22.6%) had a treatment response < 50%. Complete treatment response was significantly associated with higher concentrations of albendazole sulfoxide (P = .032), younger age (P = .032), fewer cysts (P = .049) and lower pretreatment parasite volume (P = .037). Higher number of previous cysticidal treatment courses was associated with a noncomplete treatment response (P = .023). Although the large proportion of participants with less than a complete response emphasizes the need to develop more efficacious pharmacologic regimens, the association of albendazole sulfoxide concentrations with treatment response highlights the importance of optimizing existing therapeutic regimens. In addition, the association of treatment response with parasite volume emphasizes the importance of early diagnosis.

Neurofibromatosis type 2 tumor suppressor protein is expressed in oligodendrocytes and regulates cell proliferation and process formation.

The neurofibromatosis type 2 (NF2) tumor suppressor protein Merlin functions as a negative regulator of cell growth and actin dynamics in different cell types amongst which Schwann cells have been extensively studied. In contrast, the presence and the role of Merlin in oligodendrocytes, the myelin forming cells within the CNS, have not been elucidated. In this work, we demonstrate that Merlin immunoreactivity was broadly distributed in the white matter throughout the central nervous system. Following Merlin expression during development in the cerebellum, Merlin could be detected in the cerebellar white matter tract at early postnatal stages as shown by its co-localization with Olig2-positive cells as well as in adult brain sections where it was aligned with myelin basic protein containing fibers. This suggests that Merlin is expressed in immature and mature oligodendrocytes. Expression levels of Merlin were low in oligodendrocytes as compared to astrocytes and neurons throughout development. Expression of Merlin in oligodendroglia was further supported by its identification in either immortalized cell lines of oligodendroglial origin or in primary oligodendrocyte cultures. In these cultures, the two main splice variants of Nf2 could be detected. Merlin was localized in clusters within the nuclei and in the cytoplasm. Overexpressing Merlin in oligodendrocyte cell lines strengthened reduced impedance in XCELLigence measurements and Ki67 stainings in cultures over time. In addition, the initiation and elongation of cellular projections were reduced by Merlin overexpression. Consistently, cell migration was retarded in scratch assays done on Nf2-transfected oligodendrocyte cell lines. These data suggest that Merlin actively modulates process outgrowth and migration in oligodendrocytes.

Extraparenchymal neurocysticercosis: Demographic, clinicoradiological, and inflammatory features.

BACKGROUND: Extraparenchymal neurocysticercosis (ExPNCC), an infection caused by Taenia solium cysticerci that mainly occurs in the ventricular compartment (Ve) or the basal subarachnoid space (SAb), is more severe but less frequent and much less studied than parenchymal neurocysticercosis (ParNCC). Demographic, clinical, radiological, and lumbar cerebrospinal fluid features of patients affected by ExPNCC are herein described and compared with those of ParNCC patients. METHODOLOGY AND PRINCIPAL FINDINGS: 429 patients with a confirmed diagnosis of neurocysticercosis, attending the Instituto Nacional de Neurología y Neurocirugía, a tertiary reference center in Mexico City, from 2000 through 2014, were included. Demographic information, signs and symptoms, radiological patterns, and lumbar cerebrospinal fluid (CSF) laboratory values were retrieved from medical records for all patients. Data were statistically analyzed to assess potential differences depending on cyst location and to determine the effects of age and sex on the disease presentation. In total, 238 ExPNCC and 191 ParNCC patients were included. With respect to parenchymal cysts, extraparenchymal parasites were diagnosed at an older age (P = 0.002), chiefly caused intracranial hypertension (P < 0.0001), were more frequently multiple and vesicular (P < 0.0001), and CSF from these patients showed higher protein concentration and cell count (P < 0.0001). SAb patients were diagnosed at an older age than Ve patients, and showed more frequently seizures, vesicular cysticerci, and higher CSF cellularity. Gender and age modulated some traits of the disease. CONCLUSIONS: This study evidenced clear clinical, radiological, and inflammatory differences between ExPNCC and ParNCC, and between SAb and Ve patients, and demonstrated that parasite location determines different pathological entities.

A lateral flow assay (LFA) for the rapid detection of extraparenchymal neurocysticercosis using cerebrospinal fluid.

A lateral flow assay (LFA) for the diagnosis and monitoring of extraparenchymal neurocysticercosis, has been developed. The assay is based on the use of the monoclonal antibody HP10, and when applied to cerebrospinal fluid, correctly identified 34 cases of active extraparenchymal neurocysticercosis, but was negative with 26 samples from treated and cured neurocysticercosis patients and with 20 samples from unrelated neurological diseases. There was complete agreement between the HP10 Ag-ELISA results and the HP10-LFA. The HP10-LFA thus has utility for diagnosis and treatment of extraparenchymal neurocysticercosis, frequently a more dangerous form of the infection.

Neuron-Specific Deletion of the Nf2 Tumor Suppressor Impairs Functional Nerve Regeneration.

In contrast to axons of the central nervous system (CNS), axons of the peripheral nervous system (PNS) show better, but still incomplete and often slow regeneration following injury. The tumor suppressor protein merlin, mutated in the hereditary tumor syndrome Neurofibromatosis type 2 (NF2), has recently been shown to have RhoA regulatory functions in PNS neurons-in addition to its well-characterized, growth-inhibitory activity in Schwann cells. Here we report that the conditional knockout of merlin in PNS neurons leads to impaired functional recovery of mice following sciatic nerve crush injury, in a gene-dosage dependent manner. Gross anatomical or electrophysiological alterations of sciatic nerves could not be detected. However, correlating with attenuated RhoA activation due to merlin deletion, ultrastructural analysis of nerve samples indicated enhanced sprouting of axons with reduced caliber size and increased myelination compared to wildtype animals. We conclude that deletion of the tumor suppressor merlin in the neuronal compartment of peripheral nerves results in compromised functional regeneration after injury. This mechanism could explain the clinical observation that NF2 patients suffer from higher incidences of slowly recovering facial nerve paralysis after vestibular schwannoma surgery.