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In addition to having cultural importance, village chicken production remains an important source of cash income for most rural households in Burkina Faso. However, strict biosecurity and good management of chicken flocks are required to reduce the risk of exposure of communities to chicken waste at household level. We characterized village production systems in rural Burkina Faso in relation to importance to food security, biosecurity, husbandry and chicken health management. We surveyed 483 chicken-producing households and carried out 20 focus group discussions separately with men and women chicken producers in Boussouma commune, a typical rural setting. Crop farming was reported as the main income-generating activity carried out by chicken producers (79.5%). Seveny six per cent of households sold chicken to local markets. Chicken production and sales were aligned to social/cultural events (religious festivals, weddings, etc…) and school-fee payment period. While men spent more revenues from chicken production on agriculture and household equipment's, women spent more on food, education and medical expenses. The chicken management system is mainly extensive scavenging, with most farmers (81.4%) keeping 5 to 50 birds with little or no supplementary feeding and rudimentary housing. Most producers indicated that Newcastle disease was the main cause of chicken mortality. While men consider high disease burden, lack of finance, and poor chicken housing as the major constraints, women prioritized the lack of adequate chicken housing, lack of feeds and limited access to veterinary services. With locally adapted interventions that build one a One Health approach, village chicken keeping has the potential to secure and greatly improve smallholder livelihoods and household food security, while preserving public health in Burkina Faso.

The safety of ready-to-eat food sold in urban informal markets in low and middle-income countries is a pressing public health challenge, that needs to be addressed if we are to establish healthy food systems. Guided by the Capability, Opportunity, and Motivation model for Behavior change (COM-B), this qualitative study aimed to explore perceptions of street vendors on their participation in a food safety capacity building intervention, consisting of training and provision of food safety equipment. The intervention aimed to improve food safety behavior of vendors of ready-to-eat chicken in informal markets in Ouagadougou, Burkina Faso. A total of 24 vendors selling ready-to-eat chicken at street restaurants participated in semi-structured interviews after training, which focused on vendors' stories of change related to food safety capabilities, opportunities, motivation, and behaviors. Data were thematically analyzed following COM-B components. Vendors noted improvements in psychological (i.e., knowledge, awareness, self-efficacy, perceptions) and physical capabilities (i.e., equipment useability and applicability), and motivations (perceived responsibility, reputation, client satisfaction, profits, consumer demand). Moreover, training and provision of equipment, spill-over effects to employees or neighboring outlets, and social support were perceived as key social and physical opportunities, while structural challenges such as market infrastructure, regulations, financial resources, cost of living, and outlet culture were physical barriers to implement lessons learnt. This study provides insights into the impact of engaging vendors in improving food safety behavior through training and equipment provision. Improvements in vendors' perceived capabilities and motivation contributed to improved food safety behavior, while contextual barriers hindered the perceived adoption of food safety behaviors.

ObjectiveThis study aimed to evaluate the effect of antimicrobial photodynamic therapy (aPDT) and the use of probiotics on the treatment of halitosis.MethodsFifty-two participants, aged from 18 to 25 years, exhaling sulfhydride (H2S) ≥ 112 ppb were selected. They were allocated into 4 groups (n = 13): Group 1: tongue scraper; Group 2: treated once with aPDT; Group 3: probiotic capsule containing Lactobacillus salivarius WB21 (6.7 x 108 CFU) and xylitol (280mg), 3 times a day after meals, for 14 days; Group 4: treated once with aPDT and with the probiotic capsule for 14 days. Halimetry with gas chromatography (clinical evaluation) and microbiological samples were collected from the dorsum of the tongue before and after aPDT, as well as after 7, 14, and 30 days. The clinical data failed to follow a normal distribution; therefore, comparisons were made using the Kruskal-Wallis test (independent measures) and Friedman ANOVA (dependent measures) followed by appropriate posthoc tests, when necessary. For the microbiological data, seeing as the data failed to follow a normal distribution, the Kruskal-Wallis rank sum test was performed with Dunn's post-test. The significance level was α = 0.05.ResultsClinical results (halimetry) showed an immediate significant reduction in halitosis with aPDT (p = 0.0008) and/or tongue scraper (p = 0.0006). Probiotics showed no difference in relation to the initial levels (p = 0.7530). No significant differences were found in the control appointments. The amount of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were not altered throughout the analysis (p = 0.1616, p = 0.2829 and p = 0.2882, respectively).ConclusionThere was an immediate clinical reduction of halitosis with aPDT and tongue scraping, but there was no reduction in the number of bacteria throughout the study, or differences in the control times, both in the clinical and microbiological results. New clinical trials are necessary to better assess the tested therapies.Trial registrationClinical Trials NCT03996044.

In addition to having cultural importance, village chicken production remains an important source of cash income for most rural households in Burkina Faso. However, strict biosecurity and good management of chicken flocks are required to reduce the risk of exposure of communities to chicken waste at household level. We characterized village production systems in rural Burkina Faso in relation to importance to food security, biosecurity, husbandry and chicken health management. We surveyed 483 chicken-producing households and carried out 20 focus group discussions separately with men and women chicken producers in Boussouma commune, a typical rural setting. Crop farming was reported as the main income-generating activity carried out by chicken producers (79.5%). Seveny six per cent of households sold chicken to local markets. Chicken production and sales were aligned to social/cultural events (religious festivals, weddings, etc...) and school-fee payment period. While men spent more revenues from chicken production on agriculture and household equipment's, women spent more on food, education and medical expenses. The chicken management system is mainly extensive scavenging, with most farmers (81.4%) keeping 5 to 50 birds with little or no supplementary feeding and rudimentary housing. Most producers indicated that Newcastle disease was the main cause of chicken mortality. While men consider high disease burden, lack of finance, and poor chicken housing as the major constraints, women prioritized the lack of adequate chicken housing, lack of feeds and limited access to veterinary services. With locally adapted interventions that build one a One Health approach, village chicken keeping has the potential to secure and greatly improve smallholder livelihoods and household food security, while preserving public health in Burkina Faso. [ABSTRACT FROM AUTHOR]

Anthropogenic impacts on tropical and subtropical coastal environments are increasing at an alarming rate, compromising ecosystem functions, structures and services. Understanding the scale of marine population decline and diversity loss requires a long-term perspective that incorporates information from a range of sources. The Southern Atlantic Ocean represents a major gap in our understanding of pre-industrial marine species composition. Here we begin to fill this gap by performing an extensive review of the published data on Middle and Late Holocene marine fish remains along the southern coast of Brazil. This region preserves archaeological sites that are unique archives of past socio-ecological systems and pre-European biological diversity. We assessed snapshots of species compositions and relative abundances spanning the last 9500 years, and modelled differences in species’ functional traits between archaeological and modern fisheries. We found evidence for both generalist and specialist fishing practices in pre-European times, with large body size and body mass caught regularly over hundreds of years. Comparison with modern catches revealed a significant decline in these functional traits, possibly associated with overfishing and escalating human impacts in recent times.

Vaccination is considered as the main tool for the Global Control and Eradication Strategy for peste des petits ruminants (PPR), and the efficacity of the PPR-vaccine in conferring long-life immunity has been established. Despite this, previous studies asserted that vaccination can be expensive and consequently, the effectiveness of disease control may not necessarily translate to overall profit for farmers. Also, the consequences of PPR control on socioeconomic indicators like food and nutrition security at a macro-national level have not been explored thoroughly. Therefore, this study seeks to assess ex-ante the impact of PPR control strategies on farm-level profitability and the socioeconomic consequences concerning food and nutrition security at a national level in Senegal. A bi-level system dynamics model, compartmentalised into five modules consisting of integrated production-epidemiological, economics, disease control, marketing, and policy modules, was developed with the STELLA Architect software, validated, and simulated for 30 years at a weekly timestep. The model was parameterised with data from household surveys from pastoral areas in Northern Senegal and relevant existing data. Nine vaccination scenarios were examined considering different vaccination parameters (vaccination coverage, vaccine wastage, and the provision of government subsidies). The findings indicate that compared to a no-vaccination scenario, all the vaccination scenarios for both 26.5% (actual vaccination coverage) and 70% (expected vaccination coverage) resulted in statistically significant differences in the gross margin earnings and the potential per capita consumption for the supply of mutton and goat meat. At the prevailing vaccination coverage (with or without the provision of government subsidies), farm households will earn an average gross margin of $69.43 (annually) more than without vaccination, and the average per capita consumption for mutton and goat meat will increase by 1.13kg/person/year. When the vaccination coverage is increased to the prescribed threshold for PPR eradication (i.e., 70%), with or without the provision of government subsidies, the average gross margin earnings would be $72.23 annually and the per capita consumption will increase by 1.23kg/person/year compared to the baseline (without vaccination). This study's findings offer an empirical justification for a sustainable approach to PPR eradication. The information on the socioeconomic benefits of vaccination can be promoted via sensitization campaigns to stimulate farmers' uptake of the practice. This study can inform investment in PPR control.

The safety of ready-to-eat food sold in urban informal markets in low and middle-income countries is a pressing public health challenge, that needs to be addressed if we are to establish healthy food systems. Guided by the Capability, Opportunity, and Motivation model for Behavior change (COM-B), this qualitative study aimed to explore perceptions of street vendors on their participation in a food safety capacity building intervention, consisting of training and provision of food safety equipment. The intervention aimed to improve food safety behavior of vendors of ready-to-eat chicken in informal markets in Ouagadougou, Burkina Faso. A total of 24 vendors selling ready-to-eat chicken at street restaurants participated in semi-structured interviews after training, which focused on vendors' stories of change related to food safety capabilities, opportunities, motivation, and behaviors. Data were thematically analyzed following COM-B components. Vendors noted improvements in psychological (i.e., knowledge, awareness, self-efficacy, perceptions) and physical capabilities (i.e., equipment useability and applicability), and motivations (perceived responsibility, reputation, client satisfaction, profits, consumer demand). Moreover, training and provision of equipment, spill-over effects to employees or neighboring outlets, and social support were perceived as key social and physical opportunities, while structural challenges such as market infrastructure, regulations, financial resources, cost of living, and outlet culture were physical barriers to implement lessons learnt. This study provides insights into the impact of engaging vendors in improving food safety behavior through training and equipment provision. Improvements in vendors' perceived capabilities and motivation contributed to improved food safety behavior, while contextual barriers hindered the perceived adoption of food safety behaviors. [ABSTRACT FROM AUTHOR]

Increasingly, communicating science to the public is recognized as the responsibility of professional scientists; however, these skills are not always included in graduate training. In addition, most research on science communication training during graduate school, which is limited, has been program evaluation or literature reviews and does not report on or seek to understand graduate student perspectives. This research study provides a comprehensive analysis of graduate-level science communication training from the perspective of STEM graduate students. Using a mixed-methods approach, this study aimed to investigate where graduate students are receiving science communication training (if at all), what this training looks like from the student’s point of view, and, for graduate students that are engaging in science communication, what do these experiences look like. This study also explores how graduate students define science communication. Taken together, these results will give graduate students a voice in the development of science communication trainings and will remove barriers and increase equity in science communication training.

Sickle cell anemia (SCA) is caused by a single point variation in the β-globin gene (HBB): c.20A> T (p.Glu7Val), in homozygous state. SCA is characterized by sickling of red blood cells in small blood vessels which leads to a range of multiorgan complications, including kidney dysfunction. This case-control study aims at identifying sickle cell nephropathy biomarkers in a group of patients living with SCA from Senegal. A total of 163 patients living with SCA and 177 ethnic matched controls were investigated. Biological phenotyping included evaluation of glycemia, glucosuria, albuminuria, proteinuria, tubular proteinuria, serum creatinine, urine creatinine, urine specific gravity and glomerular filtration rate. Descriptive statistics of biomarkers were performed using the χ2 –test, with the significance level set at p

Objective: This study aimed to evaluate the effect of antimicrobial photodynamic therapy (aPDT) and the use of probiotics on the treatment of halitosis. Methods: Fifty-two participants, aged from 18 to 25 years, exhaling sulfhydride (H2S) ≥ 112 ppb were selected. They were allocated into 4 groups (n = 13): Group 1: tongue scraper; Group 2: treated once with aPDT; Group 3: probiotic capsule containing Lactobacillus salivarius WB21 (6.7 x 108 CFU) and xylitol (280mg), 3 times a day after meals, for 14 days; Group 4: treated once with aPDT and with the probiotic capsule for 14 days. Halimetry with gas chromatography (clinical evaluation) and microbiological samples were collected from the dorsum of the tongue before and after aPDT, as well as after 7, 14, and 30 days. The clinical data failed to follow a normal distribution; therefore, comparisons were made using the Kruskal-Wallis test (independent measures) and Friedman ANOVA (dependent measures) followed by appropriate posthoc tests, when necessary. For the microbiological data, seeing as the data failed to follow a normal distribution, the Kruskal-Wallis rank sum test was performed with Dunn's post-test. The significance level was α = 0.05. Results: Clinical results (halimetry) showed an immediate significant reduction in halitosis with aPDT (p = 0.0008) and/or tongue scraper (p = 0.0006). Probiotics showed no difference in relation to the initial levels (p = 0.7530). No significant differences were found in the control appointments. The amount of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were not altered throughout the analysis (p = 0.1616, p = 0.2829 and p = 0.2882, respectively). Conclusion: There was an immediate clinical reduction of halitosis with aPDT and tongue scraping, but there was no reduction in the number of bacteria throughout the study, or differences in the control times, both in the clinical and microbiological results. New clinical trials are necessary to better assess the tested therapies. Trial registration: Clinical Trials NCT03996044. [ABSTRACT FROM AUTHOR]

ObjectivesThe aim of this study was to evaluate the accuracy of three consumer-based activity monitors, Fitbit Charge 2, Fitbit Alta, and the Apple Watch 2, all worn on the wrist, in estimating step counts, moderate-to-vigorous minutes (MVPA), and heart rate in a free-living setting.MethodsForty-eight participants (31 females, 17 males; ages 18-59) were asked to wear the three consumer-based monitors mentioned above on the wrist, concurrently with a Yamax pedometer as the criterion for step count, an ActiGraph GT3X+ (ActiGraph) for MVPA, and a Polar H7 chest strap for heart rate. Participants wore the monitors for a 24-hour free-living condition without changing their usual active routine. MVPA was calculated in bouts of ≥10 minutes. Pearson correlation, mean absolute percent error (MAPE), and equivalence testing were used to evaluate the measurement agreement.ResultsThe average step counts recorded for each device were as follows: 11,734 (Charge2), 11,922 (Alta), 11,550 (Apple2), and 10,906 (Yamax). The correlations in steps for the above monitors ranged from 0.84 to 0.95 and MAPE ranged from 17.1% to 35.5%. For MVPA minutes, the average were 76.3 (Charge2), 63.3 (Alta), 49.5 (Apple2), and 47.8 (ActiGraph) minutes accumulated in bouts of 10 or greater minutes. The correlation from MVPA estimation for above monitors were 0.77, 0.91, and 0.66. MAPE from MVPA estimation ranged from 44.7% to 55.4% compared to ActiGraph. For heart rate, correlation for Charge2 and Apple2 was higher for sedentary behavior and lower for MVPA. The MAPE ranged from 4% to 16%.ConclusionAll three consumer monitors estimated step counts fairly accurately, and both the Charge2 and Apple2 reported reasonable heart rate estimation. However, all monitors substantially underestimated MVPA in free-living settings.

The ecology of the early herbivorous juvenile stage of the crown-of-thorns sea star (COTS, Acanthaster spp.) is poorly understood, yet the success of this life stage is key to generating population outbreaks that devastate coral reefs. Crustose coralline algae (CCA) has been considered to be the main diet of herbivorous juveniles. In this study, we show that COTS can avail of a range of algal food. Juveniles were reared on CCA, Amphiroa sp., and biofilm, and survived for 10 months on all three diets. The juveniles fed CCA and Amphiroa sp. reached 15-16.5 mm diameter at ~ 6 months and maintained this size for the rest the experiment (an additional ~4 months). Juveniles fed biofilm grew more slowly and to a smaller maximum size (~3 mm diameter). However, when juveniles were switched from biofilm to CCA they resumed growth to a new asymptotic size (~13.5 mm, 13-20 months). In diet choice experiments, juveniles did not show a preference between Amphiroa sp. and CCA, but generally avoided biofilm. Our results show that juvenile COTS grew equally well on CCA and Amphiroa sp. and can subsist on biofilm for months. Some juveniles, mostly from the biofilm diet treatment, decreased in size for a time and this was followed by recovery. Flexibility in diet, growth, and prolonged maintenance of asymptotic size indicates capacity for growth plasticity in herbivorous juvenile COTS. There is potential for juvenile COTS to persist for longer than anticipated and increase in number as they wait for the opportunity to avail of coral prey. These findings complicate our ability to predict recruitment to the corallivorous stage and population outbreaks following larval settlement and the ability to understand the age structure of COTS populations.

Molecular approaches have long led to the assumption that the human gut microbiota is dominated by uncultivable bacteria. The recent advent of large-scale culturing methods, and in particular that of culturomics have demonstrated that these prokaryotes can in fact be cultured. This is increasing in a dramatic manner the repertoire of commensal microbes inhabiting the human gut. Following eight years of culturomics approach applied on more than 900 samples, we propose herein a remake of the pioneering study applying a dual approach including culturomics and metagenomics on a cohort of 8 healthy specimen. Here we show that culturomics enable a 20% higher richness when compared to molecular approaches by culturing 1 archaeal species and 494 bacterial species of which 19 were new taxa. Species discovered as a part of previous culturomics studies represent 30% of the cultivated isolates, while sequences derived from these new taxa enabled to increase by 22% the bacterial richness retrieved by metagenomics. Overall, 67% of the total reads generated were covered by cultured isolates, significantly reducing the hidden content of sequencing methods compared to the pioneering study. By redefining culture conditions to recover microbes previously considered fastidious, there are greater opportunities than ever to eradicate metagenomics dark matter.

Previous research on trichinellosis in Africa focused on isolating Trichinella from wildlife while the role of domestic pigs has remained highly under-researched. Pig keeping in Uganda is historically recent, and evidence on zoonotic pig diseases, including infection with Trichinella species, is scarce. A cross-sectional survey on Trichinella seroprevalence in pigs was conducted in three districts in Central and Eastern Uganda from April 2013 to January 2015. Serum from a random sample of 1125 pigs from 22 villages in Eastern and Central Uganda was examined to detect immunoglobulin G (IgG) against any Trichinella spp. using a commercially available ELISA based on excretory-secretory antigen. ELISA positive samples were confirmed using Western Blot based on somatic antigen of Trichinella spiralis as recommended in previous validation studies. Diaphragm pillar muscle samples (at least 5 g each) of 499 pigs from areas with high ELISA positivity were examined using the artificial digestion method. Overall, 78 of all 1125 animals (6.9%, 95% CI: 5.6-8.6%) tested positive for antibodies against Trichinella spp. in the ELISA at significantly higher levels in Kamuli district compared to Masaka and Mukono districts. Thirty-one percent of the ELISA positive samples were confirmed IgG positive by the Western Blot leading to an overall seroprevalence of 2.1% (95% CI: 1.4-3.2%). The large proportion of ELISA positive samples that could not be confirmed using Western blot may be the result of cross-reactivity with other gastrointestinal helminth infections or unknown host-specific immune response mechanisms in local pig breeds in Uganda. Attempts to isolate muscle larvae for species determination using the artificial digestion method were unsuccessful. Due to the large number of muscle samples examined we are confident that even if pigs are infected, the larval burden in pork is too low to pose a major risk to consumers of developing trichinellosis. This was the first large systematic field investigation of Trichinella infection in domestic pigs in Uganda and its results imply that further studies are needed to identify the Trichinella species involved, and to identify potential sources of infection for humans.

In neural structures with complex geometries, numerical resolution of the Poisson-Nernst-Planck (PNP) equations is necessary to accurately model electrodiffusion. This formalism allows one to describe ionic concentrations and the electric field (even away from the membrane) with arbitrary spatial and temporal resolution which is impossible to achieve with models relying on cable theory. However, solving the PNP equations on complex geometries involves handling intricate numerical difficulties related either to the spatial discretization, temporal discretization or the resolution of the linearized systems, often requiring large computational resources which have limited the use of this approach. In the present paper, we investigate the best ways to use the finite elements method (FEM) to solve the PNP equations on domains with discontinuous properties (such as occur at the membrane-cytoplasm interface). 1) Using a simple 2D geometry to allow comparison with analytical solution, we show that mesh adaptation is a very (if not the most) efficient way to obtain accurate solutions while limiting the computational efforts, 2) We use mesh adaptation in a 3D model of a node of Ranvier to reveal details of the solution which are nearly impossible to resolve with other modelling techniques. For instance, we exhibit a non linear distribution of the electric potential within the membrane due to the non uniform width of the myelin and investigate its impact on the spatial profile of the electric field in the Debye layer.

Spinal muscular atrophy (SMA) is an autosomal recessive motor neuron disorder. SMA is caused by homozygous loss of the SMN1 gene and retention of the SMN2 gene resulting in reduced levels of full length SMN protein that are insufficient for motor neuron function. Various treatments that restore levels of SMN are currently in clinical trials and biomarkers are needed to determine the response to treatment. Here, we sought to investigate in SMA mice a set of plasma analytes, previously identified in patients with SMA to correlate with motor function. The goal was to determine whether levels of plasma markers were altered in the SMNΔ7 mouse model of SMA and whether postnatal SMN restoration resulted in normalization of the biomarkers.SMNΔ7 and control mice were treated with antisense oligonucleotides (ASO) targeting ISS-N1 to increase SMN protein from SMN2 or scramble ASO (sham treatment) via intracerebroventricular injection on postnatal day 1 (P1). Brain, spinal cord, quadriceps muscle, and liver were analyzed for SMN protein levels at P12 and P90. Ten plasma biomarkers (a subset of biomarkers in the SMA-MAP panel available for analysis in mice) were analyzed in plasma obtained at P12, P30, and P90.Of the eight plasma biomarkers assessed, 5 were significantly changed in sham treated SMNΔ7 mice compared to control mice and were normalized in SMNΔ7 mice treated with ASO.This study defines a subset of the SMA-MAP plasma biomarker panel that is abnormal in the most commonly used mouse model of SMA. Furthermore, some of these markers are responsive to postnatal SMN restoration. These findings support continued clinical development of these potential prognostic and pharmacodynamic biomarkers.

Although previous studies suggested an anti-inflammatory property of Brazilian red propolis (BRP), the mechanisms involved in the anti-inflammatory effects of BRP and its activity on macrophages were still not elucidated. This study aimed to evaluate whether BRP attenuates the inflammatory effect of LPS on macrophages and to investigate its underlying mechanisms. BRP was added to RAW 264.7 murine macrophages after activation with LPS. NO production, cell viability, cytokines profile were evaluated. Activation of inflammatory signaling pathways and macrophage polarization were determined by RT-qPCR and Western blot. BRP at 50 μg/ml inhibited NO production by 78% without affecting cell viability. Cd80 and Cd86 were upregulated whereas mrc1 was down regulated by BRP indicating macrophage polarization at M1. BRP attenuated the production of pro-inflammatory mediators IL-12, GM-CSF, IFN-Ɣ, IL-1β in cell supernatants although levels of TNF- α and IL-6 were slightly increased after BRP treatment. Levels of IL-4, IL-10 and TGF-β were also reduced by BRP. BRP significantly reduced the up-regulation promoted by LPS of transcription of genes in inflammatory signaling (Pdk1, Pak1, Nfkb1, Mtcp1, Gsk3b, Fos and Elk1) and of Il1β and Il1f9 (fold-change rate > 5), which were further confirmed by the inhibition of NF-κB and MAPK signaling pathways. Furthermore, the upstream adaptor MyD88 adaptor-like (Mal), also known as TIRAP, involved in TLR2 and TLR4 signaling, was down- regulated in BRP treated LPS-activated macrophages. Given that BRP inhibited multiple signaling pathways in macrophages involved in the inflammatory process activated by LPS, our data indicated that BRP is a noteworthy food-source for the discovery of new bioactive compounds and a potential candidate to attenuate exhacerbated inflammatory diseases.

OBJECTIVESSpinal Muscular Atrophy (SMA) presents challenges in (i) monitoring disease activity and predicting progression, (ii) designing trials that allow rapid assessment of candidate therapies, and (iii) understanding molecular causes and consequences of the disease. Validated biomarkers of SMA motor and non-motor function would offer utility in addressing these challenges. Our objectives were (i) to discover additional markers from the Biomarkers for SMA (BforSMA) study using an immunoassay platform, and (ii) to validate the putative biomarkers in an independent cohort of SMA patients collected from a multi-site natural history study (NHS).METHODSBforSMA study plasma samples (N = 129) were analyzed by immunoassay to identify new analytes correlating to SMA motor function. These immunoassays included the strongest candidate biomarkers identified previously by chromatography. We selected 35 biomarkers to validate in an independent cohort SMA type 1, 2, and 3 samples (N = 158) from an SMA NHS. The putative biomarkers were tested for association to multiple motor scales and to pulmonary function, neurophysiology, strength, and quality of life measures. We implemented a Tobit model to predict SMA motor function scores.RESULTS12 of the 35 putative SMA biomarkers were significantly associated (pCONCLUSIONSDiscovery and validation using independent cohorts yielded a set of SMA biomarkers significantly associated with motor function and other measures of SMA disease activity. A commercial SMA-MAP biomarker panel was generated for further testing in other SMA collections and interventional trials. Future work includes evaluating the panel in other neuromuscular diseases, for pharmacodynamic responsiveness to experimental SMA therapies, and for predicting functional changes over time in SMA patients.

BACKGROUND:A village-randomized trial of a seven-valent pneumococcal-conjugate-vaccine (PCV-7) conducted in rural Gambia showed a decrease of vaccine-type (VT) and a non-significant increase in non-vaccine-type (NVT) nasopharyngeal carriage of pneumococci two years after vaccination. Here, we report findings four years after vaccination. METHODS:PCV-7 was given to all children below 30 months of age enrolled in the trial and to those born during its course in all study villages. Villages were randomized (older children and adults) to receive PCV-7 (wholly vaccinated villages) or serogroup-C-meningococcal-conjugate-vaccine (partly vaccinated villages). Cross-sectional surveys (CSS) to collect nasopharyngeal swabs were conducted before and at various intervals after vaccination. Sixteen of these randomized villages (8 wholly vaccinated and 8 partly vaccinated) participated in a CSS conducted four years after vaccination started. RESULTS:Four years after vaccination, the prevalence of VT pneumococcal carriage was slightly higher in partly than in wholly vaccinated villages [6.4% versus 3.9% (p = 0.120)] compared to 24.4% in the pre-vaccination CSS (p

Spinal Muscular Atrophy (SMA) is a neurodegenerative motor neuron disorder resulting from a homozygous mutation of the survival of motor neuron 1 (SMN1) gene. The gene product, SMN protein, functions in RNA biosynthesis in all tissues. In humans, a nearly identical gene, SMN2, rescues an otherwise lethal phenotype by producing a small amount of full-length SMN protein. SMN2 copy number inversely correlates with disease severity. Identifying other novel biomarkers could inform clinical trial design and identify novel therapeutic targets.To identify novel candidate biomarkers associated with disease severity in SMA using unbiased proteomic, metabolomic and transcriptomic approaches.A cross-sectional single evaluation was performed in 108 children with genetically confirmed SMA, aged 2-12 years, manifesting a broad range of disease severity and selected to distinguish factors associated with SMA type and present functional ability independent of age. Blood and urine specimens from these and 22 age-matched healthy controls were interrogated using proteomic, metabolomic and transcriptomic discovery platforms. Analyte associations were evaluated against a primary measure of disease severity, the Modified Hammersmith Functional Motor Scale (MHFMS) and to a number of secondary clinical measures.A total of 200 candidate biomarkers correlate with MHFMS scores: 97 plasma proteins, 59 plasma metabolites (9 amino acids, 10 free fatty acids, 12 lipids and 28 GC/MS metabolites) and 44 urine metabolites. No transcripts correlated with MHFMS.In this cross-sectional study, "BforSMA" (Biomarkers for SMA), candidate protein and metabolite markers were identified. No transcript biomarker candidates were identified. Additional mining of this rich dataset may yield important insights into relevant SMA-related pathophysiology and biological network associations. Additional prospective studies are needed to confirm these findings, demonstrate sensitivity to change with disease progression, and assess potential impact on clinical trial design.Clinicaltrials.gov NCT00756821.

BACKGROUNDThe universal presence of a gene (SMN2) nearly identical to the mutated SMN1 gene responsible for Spinal Muscular Atrophy (SMA) has proved an enticing incentive to therapeutics development. Early disappointments from putative SMN-enhancing agent clinical trials have increased interest in improving the assessment of SMN expression in blood as an early "biomarker" of treatment effect.METHODSA cross-sectional, single visit, multi-center design assessed SMN transcript and protein in 108 SMA and 22 age and gender-matched healthy control subjects, while motor function was assessed by the Modified Hammersmith Functional Motor Scale (MHFMS). Enrollment selectively targeted a broad range of SMA subjects that would permit maximum power to distinguish the relative influence of SMN2 copy number, SMA type, present motor function, and age.RESULTSSMN2 copy number and levels of full-length SMN2 transcripts correlated with SMA type, and like SMN protein levels, were lower in SMA subjects compared to controls. No measure of SMN expression correlated strongly with MHFMS. A key finding is that SMN2 copy number, levels of transcript and protein showed no correlation with each other.CONCLUSIONThis is a prospective study that uses the most advanced techniques of SMN transcript and protein measurement in a large selectively-recruited cohort of individuals with SMA. There is a relationship between measures of SMN expression in blood and SMA type, but not a strong correlation to motor function as measured by the MHFMS. Low SMN transcript and protein levels in the SMA subjects relative to controls suggest that these measures of SMN in accessible tissues may be amenable to an "early look" for target engagement in clinical trials of putative SMN-enhancing agents. Full length SMN transcript abundance may provide insight into the molecular mechanism of phenotypic variation as a function of SMN2 copy number.TRIAL REGISTRYClinicaltrials.gov NCT00756821.

OBJECTIVES: Survival Motor Neuron (SMN) protein levels may become key pharmacodynamic (PD) markers in spinal muscular atrophy (SMA) clinical trials. SMN protein in peripheral blood mononuclear cells (PBMCs) can be quantified for trials using an enzyme-linked immunosorbent assay (ELISA). We developed protocols to collect, process, store and analyze these samples in a standardized manner for SMA clinical studies, and to understand the impact of age and intraindividual variability over time on PBMC SMN signal. METHODS: Several variables affecting SMN protein signal were evaluated using an ELISA. Samples were from healthy adults, adult with respiratory infections, SMA patients, and adult SMA carriers. RESULTS: Delaying PBMCs processing by 45 min, 2 hr or 24 hr after collection or isolation allows sensitive detection of SMN levels and high cell viability (>90%). SMN levels from PBMCs isolated by EDTA tubes/Lymphoprep gradient are stable with processing delays and have greater signal compared to CPT-collected samples. SMN signal in healthy individuals varies up to 8x when collected at intervals up to 1 month. SMN signals from individuals with respiratory infections show 3-5x changes, driven largely by the CD14 fraction. SMN signal in PBMC frozen lysates are relatively stable for up to 6 months. Cross-sectional analysis of PBMCs from SMA patients and carriers suggest SMN protein levels decline with age. CONCLUSIONS: The sources of SMN signal variability in PBMCs need to be considered in the design and of SMA clinical trials, and interpreted in light of recent medical history. Improved normalization to DNA or PBMC subcellular fractions may mitigate signal variability and should be explored in SMA patients.

Genetic defects leading to the reduction of the survival motor neuron protein (SMN) are a causal factor for Spinal Muscular Atrophy (SMA). While there are a number of therapies under evaluation as potential treatments for SMA, there is a critical lack of a biomarker method for assessing efficacy of therapeutic interventions, particularly those targeting upregulation of SMN protein levels. Towards this end we have engaged in developing an immunoassay capable of accurately measuring SMN protein levels in blood, specifically in peripheral blood mononuclear cells (PBMCs), as a tool for validating SMN protein as a biomarker in SMA.A sandwich enzyme-linked immunosorbent assay (ELISA) was developed and validated for measuring SMN protein in human PBMCs and other cell lysates. Protocols for detection and extraction of SMN from transgenic SMA mouse tissues were also developed.The assay sensitivity for human SMN is 50 pg/mL. Initial analysis reveals that PBMCs yield enough SMN to analyze from blood volumes of less than 1 mL, and SMA Type I patients' PBMCs show ∼90% reduction of SMN protein compared to normal adults. The ELISA can reliably quantify SMN protein in human and mouse PBMCs and muscle, as well as brain, and spinal cord from a mouse model of severe SMA.This SMN ELISA assay enables the reliable, quantitative and rapid measurement of SMN in healthy human and SMA patient PBMCs, muscle and fibroblasts. SMN was also detected in several tissues in a mouse model of SMA, as well as in wildtype mouse tissues. This SMN ELISA has general translational applicability to both preclinical and clinical research efforts.

Objectives: Spinal Muscular Atrophy (SMA) presents challenges in (i) monitoring disease activity and predicting progression, (ii) designing trials that allow rapid assessment of candidate therapies, and (iii) understanding molecular causes and consequences of the disease. Validated biomarkers of SMA motor and non-motor function would offer utility in addressing these challenges. Our objectives were (i) to discover additional markers from the Biomarkers for SMA (BforSMA) study using an immunoassay platform, and (ii) to validate the putative biomarkers in an independent cohort of SMA patients collected from a multi-site natural history study (NHS). Methods: BforSMA study plasma samples (N = 129) were analyzed by immunoassay to identify new analytes correlating to SMA motor function. These immunoassays included the strongest candidate biomarkers identified previously by chromatography. We selected 35 biomarkers to validate in an independent cohort SMA type 1, 2, and 3 samples (N = 158) from an SMA NHS. The putative biomarkers were tested for association to multiple motor scales and to pulmonary function, neurophysiology, strength, and quality of life measures. We implemented a Tobit model to predict SMA motor function scores. Results: 12 of the 35 putative SMA biomarkers were significantly associated (p<0.05) with motor function, with a 13th analyte being nearly significant. Several other analytes associated with non-motor SMA outcome measures. From these 35 biomarkers, 27 analytes were selected for inclusion in a commercial panel (SMA-MAP) for association with motor and other functional measures. Conclusions: Discovery and validation using independent cohorts yielded a set of SMA biomarkers significantly associated with motor function and other measures of SMA disease activity. A commercial SMA-MAP biomarker panel was generated for further testing in other SMA collections and interventional trials. Future work includes evaluating the panel in other neuromuscular diseases, for pharmacodynamic responsiveness to experimental SMA therapies, and for predicting functional changes over time in SMA patients. [ABSTRACT FROM AUTHOR]

Background: Spinal Muscular Atrophy (SMA) is a neurodegenerative motor neuron disorder resulting from a homozygous mutation of the survival of motor neuron 1 (SMN1) gene. The gene product, SMN protein, functions in RNA biosynthesis in all tissues. In humans, a nearly identical gene, SMN2, rescues an otherwise lethal phenotype by producing a small amount of full-length SMN protein. SMN2 copy number inversely correlates with disease severity. Identifying other novel biomarkers could inform clinical trial design and identify novel therapeutic targets. Objective:: To identify novel candidate biomarkers associated with disease severity in SMA using unbiased proteomic, metabolomic and transcriptomic approaches. Materials and Methods:: A cross-sectional single evaluation was performed in 108 children with genetically confirmed SMA, aged 2-12 years, manifesting a broad range of disease severity and selected to distinguish factors associated with SMA type and present functional ability independent of age. Blood and urine specimens from these and 22 age-matched healthy controls were interrogated using proteomic, metabolomic and transcriptomic discovery platforms. Analyte associations were evaluated against a primary measure of disease severity, the Modified Hammersmith Functional Motor Scale (MHFMS) and to a number of secondary clinical measures. Results: A total of 200 candidate biomarkers correlate with MHFMS scores: 97 plasma proteins, 59 plasma metabolites (9 amino acids, 10 free fatty acids, 12 lipids and 28 GC/MS metabolites) and 44 urine metabolites. No transcripts correlated with MHFMS. Discussion: In this cross-sectional study, "BforSMA" (Biomarkers for SMA), candidate protein and metabolite markers were identified. No transcript biomarker candidates were identified. Additional mining of this rich dataset may yield important insights into relevant SMA-related pathophysiology and biological network associations. Additional prospective studies are needed to confirm these findings, demonstrate sensitivity to change with disease progression, and assess potential impact on clinical trial design. [ABSTRACT FROM AUTHOR]

Background: The universal presence of a gene (SMN2) nearly identical to the mutated SMN1 gene responsible for Spinal Muscular Atrophy (SMA) has proved an enticing incentive to therapeutics development. Early disappointments from putative SMN-enhancing agent clinical trials have increased interest in improving the assessment of SMN expression in blood as an early "biomarker" of treatment effect. Methods: A cross-sectional, single visit, multi-center design assessed SMN transcript and protein in 108 SMA and 22 age and gender-matched healthy control subjects, while motor function was assessed by the Modified Hammersmith Functional Motor Scale (MHFMS). Enrollment selectively targeted a broad range of SMA subjects that would permit maximum power to distinguish the relative influence of SMN2 copy number, SMA type, present motor function, and age. Results: SMN2 copy number and levels of full-length SMN2 transcripts correlated with SMA type, and like SMN protein levels, were lower in SMA subjects compared to controls. No measure of SMN expression correlated strongly with MHFMS. A key finding is that SMN2 copy number, levels of transcript and protein showed no correlation with each other. Conclusion: This is a prospective study that uses the most advanced techniques of SMN transcript and protein measurement in a large selectively-recruited cohort of individuals with SMA. There is a relationship between measures of SMN expression in blood and SMA type, but not a strong correlation to motor function as measured by the MHFMS. Low SMN transcript and protein levels in the SMA subjects relative to controls suggest that these measures of SMN in accessible tissues may be amenable to an "early look" for target engagement in clinical trials of putative SMN-enhancing agents. Full length SMN transcript abundance may provide insight into the molecular mechanism of phenotypic variation as a function of SMN2 copy number. [ABSTRACT FROM AUTHOR]