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1.
Article in English | MEDLINE | ID: mdl-38969925

ABSTRACT

The electronic health record (EHR) should contain information to support culturally responsive care and research; however, the widely used default "Asian" demographic variable in most US social systems (including EHRs) lacks information to describe the diverse experience within the Asian diaspora (e.g., ethnicities, languages). This has a downstream effect on research, identifying disparities, and addressing health equity. We were particularly interested in EHRs of autistic patients from the Asian diaspora, since the presence of a developmental diagnosis might call for culturally responsive care around understanding causes, treatments, and services to support good outcomes. The aim of this study is to determine the degree to which information about Asian ethnicity, languages, and culture is documented and accessible in the EHR, and whether it is differentially available for patients with or without autism. Using electronic and manual medical chart review, all autistic and "Asian" children (group 1; n = 52) were compared to a randomly selected comparison sample of non-autistic and "Asian" children (group 2; n = 50). Across both groups, manual chart review identified more specific approximations of racial/ethnic backgrounds in 54.5% of patients, 56% for languages spoken, and that interpretation service use was underestimated by 13 percentage points. Our preliminary results highlight that culturally responsive information was inconsistent, missing, or located in progress notes rather than a central location where it could be accessed by providers. Recommendations about the inclusion of Asian ethnicity and language data are provided to potentially enhance cultural responsiveness and support better outcomes for families with an autistic child.

3.
J Autism Dev Disord ; 2023 Apr 06.
Article in English | MEDLINE | ID: mdl-37022579

ABSTRACT

Underrepresentation of socioeconomically, culturally, and/or linguistically diverse (SCLD) children with neurodevelopmental disorders (NDD) and their families has become a focal point for researchers. This systematic review aimed to identify researchers' strategies for recruiting and retaining SCLD families of children with NDD, published between 1993 and 2018. One hundred twenty-six articles were included, and study samples were categorized as "High SCLD" and "Low SCLD". Chi-square tests of independence were used to determine associations between sample composition (i.e., High/Low SCLD sample) and study characteristics reported. Significant associations were found between sample composition and studies that explicitly stated intention to recruit SCLD families, χ2(1) = 12.70, p < .001, Phi = 0.38 (moderate); and for studies that reported the following participant characteristics: language, χ2(1) = 29.58, p < .001, Phi = 0.48 (moderate-to-large); and race/ethnicity + SES + language, χ2(1) = 19.26, p <. 001, Phi = 0.39 (moderate). However, associations were not found between recruitment and retention approaches and whether studies included High SCLD or Low SCLD samples. Further study of NDD researchers' recruitment and retention approaches that successfully include SCLD families is needed.

4.
Autism ; 27(4): 997-1010, 2023 05.
Article in English | MEDLINE | ID: mdl-36510836

ABSTRACT

LAY ABSTRACT: Raising an autistic child can affect many aspects of families' lives. Parents are responsible for many decisions, from initiating evaluation to selecting and implementing treatments. How parents conceptualize the course and nature of their child's diagnosis influences these processes and parents' own well-being. Parents' perceptions about their children's autism are also affected by cultural contexts and understanding of autism. The Illness Perception Questionnaire-Revised (IPQ-R) is widely used to study cognitions in chronic health research and has been adapted and validated to measure parents' perceptions and beliefs about their children's ASD (IPQ-R-ASD). However, such studies are mostly conducted in high-income countries (HICs) with western, individualistic cultural values (e.g. United States, Canada). Therefore, it is unclear whether the IPQ-R-ASD is a useful instrument in understanding parents' perceptions of autism in Vietnam, a lower- and middle-income country (LMIC) with collectivistic Asian cultural values. These differences suggest that parents in Vietnam may have cognitive representations of their children's autism that differ from those of parents living in HIC, western countries. The purpose of this study was to examine the usability of the translated Vietnamese IPQ-R-ASD that may, ultimately, help explore Vietnamese parents' autism perceptions. While the study's result indicated the usability of the translated measure in Vietnam, when interpreted with Vietnamese norms, results also highlighted notable differences between Vietnamese and North American parents' perceptions of autism that warrant further research.


Subject(s)
Autism Spectrum Disorder , Autistic Disorder , Child , Humans , United States , Cross-Cultural Comparison , Vietnam , Autism Spectrum Disorder/psychology , Parents/psychology
5.
Paediatr Drugs ; 22(1): 95-104, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31960360

ABSTRACT

AIM: The main burden of hypoxic-ischemic encephalopathy falls in low-income countries. 2-Iminobiotin, a selective inhibitor of neuronal and inducible nitric oxide synthase, has been shown to be safe and effective in preclinical studies of birth asphyxia. Recently, safety and pharmacokinetics of 2-iminobiotin treatment on top of hypothermia has been described. Since logistics and the standard of medical care are very different in low-resource settings, the aim of this study was to investigate safety and pharmacokinetics of Two-IminoBiotin in the Democratic Republic of Congo (TIBC). METHODS: Near-term neonates, born in Kinshasa, Democratic Republic of Congo, with a Thompson score ≥ 7 were eligible for inclusion. Excluded were patients with (1) inability to insert an umbilical venous catheter for administration of the study drug; (2) major congenital or chromosomal abnormalities; (3) birth weight < 1800 g; (4) clear signs of infection; and (5) moribund patients. Neonates received six infusions of 2-iminobiotin 0.16 mg/kg started within 6 h after birth, with 4-h intervals, targeting an AUC0-4h of 365 ng*h/mL. Safety, defined as vital signs, the need for clinical intervention after administration of study drug, occurrence of (serious) adverse events, and pharmacokinetics were assessed. RESULTS: After parental consent, seven patients were included with a median Thompson score of 10 (range 8-16). No relevant changes in vital signs were observed over time. There was no need for clinical intervention due to administration of study drug. Three patients died, two after completing the study protocol, one was moribund at inclusion and should not have been included. Pharmacokinetic data of 2-iminobiotin were best described using a two-compartment model. Median AUC0-4h was 664 ng*h/mL (range 414-917). No safety issues attributed to the administration of 2-iminobiotin were found. CONCLUSION: The present dosing regimen resulted in higher AUCs than targeted, necessitating a change in the dose regimen in future efficacy trials. No adverse effects that could be attributed to the use of 2-iminobiotin were observed. EudraCT number 2015-003063-12.


Subject(s)
Asphyxia/drug therapy , Biotin/analogs & derivatives , Adult , Biotin/pharmacology , Biotin/therapeutic use , Female , Humans , Infant, Newborn , Male , Poverty
6.
J Chromatogr A ; 1584: 115-125, 2019 Jan 11.
Article in English | MEDLINE | ID: mdl-30473111

ABSTRACT

Food allergy is a growing health problem worldwide; thus, there is an urgent need for robust, specific, and sensitive analytical methods for detecting allergens. Mass spectrometry is an alternative to the existing methods, and it can overcome their limitations. One of the first steps in the development of any analytical method is the identification of the analytes to be further studied. In the case of allergen detection by mass spectrometry, the analytes are peptides. In this study, a strategy was developed for identifying potential peptide biomarkers in processed food products. This strategy was applied to processed egg matrices, and 16 potential peptide biomarkers were identified for the further detection and quantification of egg by means of mass spectrometry. With an empirical approach based on dedicated sample preparation, including tandem Lys-C/trypsin enzymatic digestion and high-resolution mass spectrometry analysis, hundreds of peptides from egg proteins were identified. This list of peptides was further refined with a series of criteria, obtained from empirical evidence, to identify the ideal biomarkers for the development of a quantitative method. These criteria include the resistance to food processing and the specificity of the peptides for eggs but also the effects of amino acid modifications and enzymatic digestion efficiency.


Subject(s)
Allergens/analysis , Biomarkers/analysis , Egg Proteins/analysis , Eggs/analysis , Food Contamination/analysis , Peptide Fragments/analysis , Tandem Mass Spectrometry/methods , Allergens/chemistry , Animals , Biomarkers/chemistry , Chickens , Egg Hypersensitivity/immunology , Egg Hypersensitivity/prevention & control , Egg Proteins/immunology , Food Handling , Humans , Peptide Fragments/immunology
7.
Food Chem ; 274: 35-45, 2019 Feb 15.
Article in English | MEDLINE | ID: mdl-30372950

ABSTRACT

Worldwide, mass spectrometry is widely used to detect and quantify food allergens, especially in complex and processed food products. Yet, the absence of a regulatory framework for the developed methods has led to a lack of harmonization between laboratories. In this study, ten allergens were analyzed in eight food products by UHPLC-MS/MS, in order to establish criteria for the retention time, variation tolerance, the ion ratio deviation, and the signal-to-noise ratio for allergen detection. The set of criteria should help laboratories to compare results and avoid false positives and negatives. Furthermore, a strategy combining standard addition and labeled peptide correction was used to quantify milk, soy, peanut, and egg allergens in eight food products. This strategy is particularly interesting for routine laboratories, which receive hundreds of samples and cannot use an external calibration curve for each sample.


Subject(s)
Allergens/analysis , Food Analysis/methods , Peptides/analysis , Tandem Mass Spectrometry/methods , Animals , Arachis/chemistry , Calibration , Chromatography, High Pressure Liquid/methods , Egg Hypersensitivity , Eggs/analysis , Food Analysis/standards , Food Hypersensitivity , Humans , Laboratories , Milk/chemistry , Reproducibility of Results , Signal-To-Noise Ratio , Tandem Mass Spectrometry/standards
8.
Food Chem ; 245: 981-988, 2018 Apr 15.
Article in English | MEDLINE | ID: mdl-29287469

ABSTRACT

Feed sustainability is one of the biggest challenges for the next few years. Solutions have to be found that take feed quality and safety into account. Animal by-products are one valuable source of proteins. However, since the bovine spongiform encephalopathy (BSE) crisis, their use has been strictly regulated. The objective of this study was to propose a routine, sensitive and specific method using ultra-high performance liquid chromatography coupled to tandem mass spectrometry for the detection of blood-derived products and milk powder in feed. Contaminated aquafeeds were analysed in order to evaluate the sensitivity and specificity of the method. This new method meets both selectivity and sensitivity (0.1% (w/w)) requirements imposed by the European Commission for animal proteins detection methods. It offers an innovative and complementary solution for the simultaneously identification of authorised and unauthorised animal by-products such as processed animal proteins (PAPs).


Subject(s)
Animal Feed/analysis , Blood , Limit of Detection , Milk/chemistry , Tandem Mass Spectrometry/methods , Animals , Cattle , Chromatography, High Pressure Liquid , Food Contamination/analysis , Time Factors
9.
J Chromatogr A ; 1530: 138-151, 2017 Dec 29.
Article in English | MEDLINE | ID: mdl-29169644

ABSTRACT

Food allergy is a considerable heath problem, as undesirable contaminations by allergens during food production are still widespread and may be dangerous for human health. To protect the population, laboratories need to develop reliable analytical methods in order to detect allergens in various food products. Currently, a large majority of allergen-related food recalls concern bakery products. It is therefore essential to detect allergens in unprocessed and processed foodstuffs. In this study, we developed a method for detecting ten allergens in complex (chocolate, ice cream) and processed (cookie, sauce) foodstuffs, based on ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Using a single protocol and considering a signal-to-noise ratio higher than 10 for the most abundant multiple reaction monitoring (MRM) transition, we were able to detect target allergens at 0.5mg/kg for milk proteins, 2.5mg/kg for peanut, hazelnut, pistachio, and cashew proteins, 3mg/kg for egg proteins, and 5mg/kg for soy, almond, walnut, and pecan proteins. The ability of the method to detect 10 allergens with a single protocol in complex and incurred food products makes it an attractive alternative to the ELISA method for routine laboratories.


Subject(s)
Allergens/analysis , Chromatography, High Pressure Liquid/methods , Food Analysis/methods , Tandem Mass Spectrometry , Chocolate/analysis , Egg Proteins/analysis , Enzyme-Linked Immunosorbent Assay , Food Hypersensitivity , Ice Cream/analysis , Milk Proteins/analysis , Nuts/chemistry , Signal-To-Noise Ratio
10.
J Chromatogr A ; 1464: 115-23, 2016 Sep 16.
Article in English | MEDLINE | ID: mdl-27554027

ABSTRACT

Sensitive detection of food allergens is affected by food processing and foodstuff complexity. It is therefore a challenge to detect cross-contamination in food production that could endanger an allergic customer's life. Here we used ultra-high performance liquid chromatography coupled to tandem mass spectrometry for simultaneous detection of traces of milk (casein, whey protein), egg (yolk, white), soybean, and peanut allergens in different complex and/or heat-processed foodstuffs. The method is based on a single protocol (extraction, trypsin digestion, and purification) applicable to the different tested foodstuffs: chocolate, ice cream, tomato sauce, and processed cookies. The determined limits of quantitation, expressed in total milk, egg, peanut, or soy proteins (and not soluble proteins) per kilogram of food, are: 0.5mg/kg for milk (detection of caseins), 5mg/kg for milk (detection of whey), 2.5mg/kg for peanut, 5mg/kg for soy, 3.4mg/kg for egg (detection of egg white), and 30.8mg/kg for egg (detection of egg yolk). The main advantage is the ability of the method to detect four major food allergens simultaneously in processed and complex matrices with very high sensitivity and specificity.


Subject(s)
Allergens/chemistry , Chromatography, High Pressure Liquid/methods , Food Analysis/methods , Food Contamination/analysis , Tandem Mass Spectrometry/methods , Animals , Arachis/chemistry , Arachis/immunology , Chickens , Eggs , Food Handling , Milk/chemistry , Milk/immunology , Soybean Proteins/chemistry , Soybean Proteins/immunology
11.
Food Chem ; 213: 417-424, 2016 Dec 15.
Article in English | MEDLINE | ID: mdl-27451199

ABSTRACT

Animal by-products are valuable protein sources in animal nutrition. Among them are blood products and blood meal, which are used as high-quality material for their beneficial effects on growth and health. Within the framework of the feed ban relaxation, the development of complementary methods in order to refine the identification of processed animal proteins remains challenging. The aim of this study was to identify specific biomarkers that would allow the detection of bovine blood products and processed animal proteins using tandem mass spectrometry. Seventeen biomarkers were identified: nine peptides for bovine plasma powder; seven peptides for bovine haemoglobin powder, including six peptides for bovine blood meal; and one peptide for porcine blood. They were not detected in several commercial compound feed or feed materials, such as blood by-products of other animal origins, milk-derived products and fish meal. These biomarkers could be used for developing a species-specific and blood-specific detection method.


Subject(s)
Animal Feed/analysis , Blood Proteins/analysis , Spectrometry, Mass, Electrospray Ionization/methods , Tandem Mass Spectrometry/methods , Amino Acid Sequence , Animals , Biomarkers/analysis , Biomarkers/blood , Blood Proteins/genetics , Cattle , Chromatography, High Pressure Liquid/methods , Dairy Products/analysis , Encephalopathy, Bovine Spongiform/diagnosis , Encephalopathy, Bovine Spongiform/prevention & control , Food Contamination/analysis , Hemoglobins/analysis , Hemoglobins/genetics , Poultry , Species Specificity , Swine
12.
J Proteomics ; 147: 125-131, 2016 09 16.
Article in English | MEDLINE | ID: mdl-27268957

ABSTRACT

UNLABELLED: The rapidly growing aquaculture industry drives the search for sustainable protein sources in fish feed. In the European Union (EU) since 2013 non-ruminant processed animal proteins (PAP) are again permitted to be used in aquafeeds. To ensure that commercial fish feeds do not contain PAP from prohibited species, EU reference methods were established. However, due to the heterogeneous and complex nature of PAP complementary methods are required to guarantee the safe use of this fish feed ingredient. In addition, there is a need for tissue specific PAP detection to identify the sources (i.e. bovine carcass, blood, or meat) of illegal PAP use. In the present study, we investigated and compared different protein extraction, solubilisation and digestion protocols on different proteomics platforms for the detection and differentiation of prohibited PAP. In addition, we assessed if tissue specific PAP detection was feasible using proteomics tools. All work was performed independently in two different laboratories. We found that irrespective of sample preparation gel-based proteomics tools were inappropriate when working with PAP. Gel-free shotgun proteomics approaches in combination with direct spectral comparison were able to provide quality species and tissue specific data to complement and refine current methods of PAP detection and identification. SIGNIFICANCE: To guarantee the safe use of processed animal protein (PAP) in aquafeeds efficient PAP detection and monitoring tools are required. The present study investigated and compared various proteomics workflows and shows that the application of shotgun proteomics in combination with direct comparison of spectral libraries provides for the desired species and tissue specific classification of this heat sterilized and pressure treated (≥133°C, at 3bar for 20min) protein feed ingredient.


Subject(s)
Animal Feed/analysis , Proteins/analysis , Proteomics/methods , Animal Feed/standards , Animals , Aquaculture/methods , Cattle , Fisheries , Fishes , Species Specificity
13.
PLoS One ; 9(5): e97131, 2014.
Article in English | MEDLINE | ID: mdl-24824620

ABSTRACT

INTRODUCTION: Churches occupy an important social and cultural position in Madagascar. The sexual transmission of HIV raises controversies about the role that Churches can play in preventing HIV/AIDS. This cross-sectional survey investigated recommendations by religious leaders for condom use and other preventive strategies in the context of international guidelines. METHODS: A questionnaire was self-administered to a random sample of religious leaders. The questions related to preventive methods against HIV/AIDS such as: condom use, marital fidelity, sexual abstinence before marriage, and HIV-testing. Associations with recommendations for condom use were evaluated using univariate and multivariate logistic regression analyses. RESULTS: Of 231 religious leaders, 215 (93.1%) were willing to share their knowledge of HIV/AIDS with their congregations. The majority received their information from the media (N=136, 58.9%), a minority from their church (N=9, 3.9%), and 38 (16.4%) had received prior training on HIV. Nearly all (N=212, 91.8%) knew that HIV could be sexually transmitted though only a few (N=39, 16.9%) were aware of mother-to-child transmission or unsafe injections (N=56, 24.2%). A total of 91 (39.4%) were willing to, or had recommended (N=64, 27.7%), condom use, while 50 (21.6%) had undergone HIV testing. Only nine (3.9%) had ever cared for a person living with HIV/AIDS (PLHIV). Multivariable logistic regression shows that condom use recommendations by religious leaders were negatively associated with tertiary level education (OR: 0.3, 95% CI 0.1-0.7), and positively associated with knowing a person at risk (OR: 16.2, 95% CI 3.2-80.2), knowing of an ART center (OR: 2.6, 95% CI 1.4-4.8), and receiving information about HIV at school (OR: 2.6, 95% CI 1.2-5.6). CONCLUSIONS: Malagasy church leaders could potentially become key players in HIV/AIDS prevention if they improved their knowledge of the illness, their commitment to international recommendations, and extended their interaction with people most at risk.


Subject(s)
Acquired Immunodeficiency Syndrome/epidemiology , Acquired Immunodeficiency Syndrome/prevention & control , Epidemics/prevention & control , Religion and Medicine , Religion and Sex , Religious Personnel/psychology , Christianity , Cross-Sectional Studies , Health Education , Health Knowledge, Attitudes, Practice , Humans , Logistic Models , Madagascar/epidemiology , Religious Personnel/statistics & numerical data , Surveys and Questionnaires
14.
Article in English | MEDLINE | ID: mdl-24674818

ABSTRACT

Domestication might be a possible way to reduce the physiological response to long-term stressors and deleterious effects on immunity. The present study aimed to evaluate the chronic immune response induced by repeated emersions and the possible impact of domestication by comparing farmed Eurasian perch with short (F1) and long (F4) captive-life history. In the first experiment, fish were exposed to a single emersion and physiological stress response was measured in the short term to characterize fish sensitivity to the tested stressor. Serum cortisol and glucose elevated within 6h post-stress and splenosomatic index (SSI) decreased within 48h, indicating that the species was affected by emersion stressor. In the second experiment, F1 and F4 generations were submitted to repeated water emersions (3 times/week during 44days). On day 9, 18 and 44, samplings were performed 48h post-stressor to highlight any sustained disruption of immune system. Serum cortisol, glucose, SSI and lysozyme activity were evaluated and serum proteome was analyzed using 2D-DIGE. Any of the tested variables were affected by repeated emersions and proteomic analysis only revealed that alpha-2 macroglobulins (a2Ms) were up-regulated in the serum of stressed individuals. Domestication also resulted in the up-regulation of five a2M isoforms and down-regulation of complement C3 and Ig light chain proteins, independently of any stressor exposure. In conclusion, the results suggested that repeated emersions are not severe stressors for Eurasian perch, probably explaining why domestication had no influence on fish responses. Changes associated with domestication are highly complex and certainly need further investigations.

15.
J Proteomics ; 94: 89-109, 2013 Dec 06.
Article in English | MEDLINE | ID: mdl-24060998

ABSTRACT

The identification of the regulatory proteins that control DNA transcription as well as RNA stability and translation represents a key step in the comprehension of gene expression regulation. Those proteins can be purified by DNA- or RNA-affinity chromatography, followed by identification by mass spectrometry. Although very simple in the concept, this represents a real technological challenge due to the low abundance of regulatory proteins compared to the highly abundant proteins binding to nucleic acids in a nonsequence-specific manner. Here we review the different strategies that have been set up to reach this purpose, discussing the key parameters that should be considered to increase the chances of success. Typically, two categories of biological questions can be distinguished: the identification of proteins that specifically interact with a precisely defined binding site, mostly addressed by quantitative mass spectrometry, and the identification in a non-comparative manner of the protein complexes recruited by a poorly characterized long regulatory region of nucleic acids. Finally, beside the numerous studies devoted to in vitro-assembled nucleic acid-protein complexes, the scarce data reported on proteomic analyses of in vivo-assembled complexes are described, with a special emphasis on the associated challenges.


Subject(s)
Chromatography, Affinity/methods , DNA-Binding Proteins , DNA/chemistry , Mass Spectrometry/methods , RNA-Binding Proteins , RNA/chemistry , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/isolation & purification , RNA-Binding Proteins/chemistry , RNA-Binding Proteins/isolation & purification
16.
Nucleic Acids Res ; 40(21): e168, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22904091

ABSTRACT

To depict the largest picture of a core promoter interactome, we developed a one-step DNA-affinity capture method coupled with an improved mass spectrometry analysis process focused on the identification of low abundance proteins. As a proof of concept, this method was developed through the analysis of 230 bp contained in the 5'long terminal repeat (LTR) of the human immunodeficiency virus 1 (HIV-1). Beside many expected interactions, many new transcriptional regulators were identified, either transcription factors (TFs) or co-regulators, which interact directly or indirectly with the HIV-1 5'LTR. Among them, the homeodomain-containing TF myeloid ectopic viral integration site was confirmed to functionally interact with a specific binding site in the HIV-1 5'LTR and to act as a transcriptional repressor, probably through recruitment of the repressive Sin3A complex. This powerful and validated DNA-affinity approach could also be used as an efficient screening tool to identify a large set of proteins that physically interact, directly or indirectly, with a DNA sequence of interest. Combined with an in silico analysis of the DNA sequence of interest, this approach provides a powerful approach to select the interacting candidates to validate functionally by classical approaches.


Subject(s)
HIV Long Terminal Repeat , HIV-1/genetics , Homeodomain Proteins/metabolism , Neoplasm Proteins/metabolism , Proteomics/methods , Transcription Factors/metabolism , Binding Sites , DNA-Binding Proteins/analysis , DNA-Binding Proteins/isolation & purification , HeLa Cells , Homeodomain Proteins/physiology , Humans , Mass Spectrometry , Myeloid Ecotropic Viral Integration Site 1 Protein , NF-kappa B/analysis , Neoplasm Proteins/physiology , Repressor Proteins/analysis , Transcription, Genetic
17.
Theriogenology ; 78(2): 432-42, 442.e1-13, 2012 Jul 15.
Article in English | MEDLINE | ID: mdl-22578620

ABSTRACT

The success of reproduction depends greatly upon gamete quality, especially oocytes which carry most of the molecular material necessary for early embryogenesis. However, it remains difficult to find relevant morphologic and/or biochemical parameters to assess oocyte quality and thus have a reliable prediction of the reproduction performance. To understand which criteria are the most reliable to assess the reproductive success of the Eurasian perch (Perca fluviatilis), we measured 14 parameters characterizing female, spawn, oocyte, and embryonic or larval development on 20 independent spawn. A data analysis allowed the definition of two clusters of spawn with different larval characteristics: the first cluster was composed of spawn which led mainly to strong large larvae presenting a low deformity rate, while the second cluster rather corresponds to spawn leading to smaller and weaker larvae with a higher deformity rate. Moreover, a third cluster (unfertilized spawn) was studied. Our analysis revealed that most of the prefertilization biological traits that we studied appeared poorly relevant to predict larval features, proper embryonic development and deformity occurrences. We thus performed a large scale proteomic analysis to highlight proteins differently expressed in each spawn cluster. A 2D-DIGE study followed by an MS/MS spectrometry allowed the identification of 32 proteins involved in several biological functions and differently expressed between spawn clusters. Among them, proteins involved in cell response to the oxidative stress, as well as energetic metabolism, heat shock proteins and Vitellogenins are of particular interest. Several functions appear specific to a spawn cluster and could thus explain their corresponding reproduction performance. In the future, proteins involved in those cellular mechanisms may constitute molecular markers predictive of the reproduction performance in Perca fluviatilis.


Subject(s)
Gene Expression Profiling/veterinary , Gene Expression Regulation, Developmental/physiology , Oocytes/metabolism , Perches/physiology , Animals , Electrophoresis, Gel, Two-Dimensional/veterinary , Female , Fish Proteins/genetics , Fish Proteins/metabolism , Principal Component Analysis , Proteomics , Reproduction
18.
Fish Shellfish Immunol ; 31(6): 1113-21, 2011 Dec.
Article in English | MEDLINE | ID: mdl-22008286

ABSTRACT

The current study aimed to evaluate the influence of domestication process on the stress response and subsequent immune modulation in Eurasian perch juveniles (Perca fluviatilis) submitted to chronic confinement. Briefly, F1 and F4 generations were confined into small-size tanks and sampled 7 and 55 days after stocking. Cortisol and glucose levels as well as lysozyme activity and immunoglobulin level were evaluated in the serum. Spleen Somatic Index and spleen ROS production were also measured. A proteomic analysis was performed on serum sampled on day 7. Finally, both generations were genetically characterized using a microsatellite approach. Globally, results revealed that chronic confinement did not elicit a typical stress response but resulted in a prolonged immune stimulation. Proteomic results suggested that domestication process influenced the immune status of perch submitted to chronic confinement as the F1 confined fish displayed lower abundance of C3 complement component, transferrin and Apolipoprotein E. Microsatellite data showed a strong genetic drift as well as reduced genetic diversity, allelic number and heterozygosity along with domestication process. The present work is the first to report that fish under domestication can develop an immune response, assessed by a combined approach, following recurrent challenges imposed by captive environment despite a reduced genetic variation.


Subject(s)
Animals, Domestic/immunology , Aquaculture/methods , Confined Spaces , Genetic Variation , Immunomodulation/immunology , Perches/immunology , Stress, Physiological/immunology , Animals , Animals, Domestic/blood , Animals, Domestic/genetics , Apolipoproteins E/immunology , Blood Glucose/analysis , Complement C3/immunology , Hydrocortisone/blood , Immunoglobulins/blood , Microsatellite Repeats/genetics , Muramidase/blood , Muramidase/immunology , Perches/blood , Perches/genetics , Reactive Oxygen Species/metabolism , Spleen/metabolism , Transferrin/immunology
19.
Biochimie ; 88(10): 1409-17, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16765502

ABSTRACT

This work shows that the proximal promoter of the mouse Afp gene contains a Ku binding site and that Ku binding is associated with down-regulation of the transcriptional activity of the Afp promoter. The Ku binding site is located in a segment able to adopt a peculiar structured form, probably a hairpin structure. Interestingly, the structured form eliminates the binding sites of the positive transcription factor HNF1. Furthermore, a DNAse hypersensitive site is detected in footprinting experiments done with extracts of AFP non-expressing hepatoma cells. These observations suggest that the structured form is stabilised by Ku and is associated with extinction of the gene in AFP non-expressing hepatic cells.


Subject(s)
Antigens, Nuclear/metabolism , DNA-Binding Proteins/metabolism , DNA/chemistry , Gene Expression Regulation , Promoter Regions, Genetic , alpha-Fetoproteins/genetics , Animals , Base Sequence , Binding Sites , Cell Line, Tumor , DNA/metabolism , Hepatocyte Nuclear Factor 1 , Humans , Ku Autoantigen , Mice , Molecular Sequence Data , Nucleic Acid Conformation , Rats , alpha-Fetoproteins/metabolism
20.
J Inorg Biochem ; 86(2-3): 547-54, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11566326

ABSTRACT

Previous investigations of the potential of metal-organic compounds as inhibitors of human immunodeficiency virus type I protease (HIV-1 PR) showed that the copper(II) complex diaqua [bis(2-pyridylcarbonyl)amido] copper(II) nitrate dihydrate and the complex bis[N2-(2,3,6-trimethoxybenzyl)-4-2-pyridinecarboxamide] copper(II) behaved as inhibitors of HIV-1 PR. In a search for similar readily accessible ligands, we synthesised and studied the structural properties of N2-(2-pyridylmethyl)-2-pyridinecarboxamide (L) copper(II) complexes. Three different crystal structures were obtained. Two were found to contain ligand L simultaneously in a tridentate and bidentate conformation [Cu(L(tri)L(bi))]. The other contained two symmetry-related ligands, coordinated through the pyridine nitrogen and the amide oxygen atoms [Cu(L(bi))(2)]. A search of the Cambridge Structural Database indicated that L(tri) resulting from nitrogen bound amide hydrogen metal substitution is favoured over chelation through the amide oxygen atom. In our case, we calculated that the conformation of L(tri) is 11 kcal/mol more favourable than that of L(bi). ESI-MS experiments showed that the Cu(L(bi))(2) structure could not be observed in solution, while Cu(L(tri)L(bi))-related complexes were indeed present. The lack of protease inhibition of the pyridine carboxamide copper(II) complexes was explained by the fact that the Cu(L(bi)L(tri)) complex could not fit into the HIV-1 active site.


Subject(s)
Copper/chemistry , HIV Protease Inhibitors/chemistry , HIV Protease Inhibitors/chemical synthesis , Organometallic Compounds/chemistry , Organometallic Compounds/chemical synthesis , Pyridines/chemistry , Pyridines/chemical synthesis , Catalytic Domain , Crystallography, X-Ray , Drug Design , HIV Protease/chemistry , HIV Protease Inhibitors/pharmacology , In Vitro Techniques , Models, Molecular , Organometallic Compounds/pharmacology , Pyridines/pharmacology , Spectrometry, Mass, Electrospray Ionization
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