Bio-detoxification of mycotoxins by lactic acid bacteria from different food matrices

Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP- PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 g/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 m pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Uncovering the role of misfolded SOD1 in the pathogenesis of Amyotrophic Lateral Sclerosis

La sclérose latérale amyothrophique (SLA) est une maladie neurodégénérative charactérisée par la perte des neurones moteurs menant à la paralysie et à la mort. Environ 20% des cas familiaux de la SLA sont causés par des mutations de la superoxyde dismutase 1 (SOD1), conduisant vers un mauvais repliement de la protéine SOD1, ce qui a comme conséquence un gain de fonction toxique. Plusieurs anticorps spécifiques pour la forme mal repliée de la protéine ont été générés et utilisés comme agent thérapeutique dans des modèles précliniques. Comment le mauvais repliement de SOD1 provoque la perte sélective des neurones moteurs demeure non résolu. La morphologie, le bilan énergétique et le transport mitochondrial sont tous documentés dans les modèles de la SLA basés sur SOD1, la détérioration des mitochondries joue un rôle clé dans la dégénération des neurones moteurs. De plus, la protéine SOD1 mal repliée s’associe sélectivement sur la surface des mitochondries de la moelle épinière chez les modèles de rongeurs de la SLA. Notre hypothèse est que l’accumulation de la protéine SOD1 mal repliée sur les mitochondries pourrait nuire aux fonctions mitochondriales. À cette fin, nous avons développé un nouvel essai par cytométrie de flux afin d’isoler les mitochondries immunomarquées avec des anticorps spécifiques à la forme malrepliée de SOD1 tout en évaluant des aspects de la fonction mitochondriale. Cette méthode permettra de comparer les mitochondries portant la protéine SOD1 mal repliée à celles qui ne la portent pas. Nous avons utilisé un anticorps à conformation spécifique de SOD1, B8H10, pour démontrer que la protéine mal repliée SOD1 s’associe avec les mitochondries de la moelle épinière des rat SOD1G93A d’une manière dépendante du temps. Les mitochondries avec la protéine mal repliée SOD1 B8H10 associée à leur surface (B8H10+) ont un volume et une production excessive de superoxyde significativement plus grand, mais possèdent un potentiel transmembranaire comparable aux mitochondries B8H10-. En outre, la présence de la protéine mal repliée SOD1 reconnue par B8H10 coïncide avec des niveaux plus élevés de la forme pro-apoptotique de Bcl-2. L’immunofluorescence de sections de moelle épinière du niveau lombaire avec l’anticorps spécifique à la conformation B8H10 et AMF7-63, un autre anticorps conformationnel spécifique de SOD1, démontre des motifs de localisations distincts. B8H10 a été trouvé principalement dans les neurones moteurs et dans plusieurs points lacrymaux dans tout le neuropile. Inversement, AMF7-63 a marqué les neurones moteurs ainsi qu’un réseau fibrillaire distinctif concentré dans la corne antérieure. Au niveau subcellulaire, SOD1 possèdant la conformation reconnu par AMF7-63 est aussi localisée sur la surface des mitochondries de la moelle épinière d’une manière dépendante du temps. Les mitochondries AMF7-63+ ont une augmentation du volume comparé aux mitochondries B8H10+ et à la sous-population non marquée. Cependant, elles produisent une quantité similaire de superoxyde. Ensemble, ces données suggèrent qu’il y a plusieurs types de protéines SOD1 mal repliées qui convergent vers les mitochondries et causent des dommages. De plus, différentes conformations de SOD1 apportent une toxicité variable vers les mitochondries. Les protéines SOD1 mal repliées réagissant à B8H10 et AMF7-63 sont présentes en agrégats dans les fractions mitochondriales, nous ne pouvons donc pas prendre en compte leurs différents effets sur le volume mitochondrial. Les anticorps conformationnels sont des outils précieux pour identifier et caractériser le continuum du mauvais repliement de SOD1 en ce qui concerne les caractéristiques biochimiques et la toxicité. Les informations présentes dans cette thèse seront utilisées pour déterminer le potentiel thérapeutique de ces anticorps.

Packaging materials for shrimp, fish and fish products, their properties, selection and effect of different packaging materials on their shelf life

Packaging is important not only in extending the shellife of fish and fishery products but also improving their marketability. In the recent years, significant development have taken place in the packaging industry. During the past decade in India, there is almost a packaging revolution with the availability of variety packaging materials, thus generating better packaging consciousness in other producer/manufacturing industries. But unfortunately, such realisation is not forthcoming in the fisheries sector and packaging techniques for local and export trade continues to be on traditional lines with their inherent drawbacks and limitations. Better packaging ensures improved quality and presentation of the products and ensures higher returns to the producer. Among several packaging materials used in fishery industry, ISI specifications had been formulated only for corrugated fibre board boxes for export of seafoods and froglegs. This standard was formulated before containersiation came into existance in the export of marine products. Before containerisation, the standards were stringent in view of the rough handling, transportation and storage. Two of the common defects reported in the master cartons exported from India are low mechanical strength and tendency to get wet. They are weakened by the deposits of moisture caused by temperature fluctuations during loading, unloading and other handling stages. It is necessary to rectify the above defects in packaging aquatic products and hence in the present study extensive investigations are carried out to find out the reasons for the damage of master cartons, to evolve code of practice for the packaging oi frozen shrimp for exports, development of alternative style of packaging for the shipping container, development of suitable consumer packaging materials for fish soup powder, cured dried mackeral, fish pickles in oil and frozen shrimp. For the development of suitable packaging materials, it is absolutely essential to know the properties of packaging materials, effect of different packaging materials on theirshelf life and their suitability for food contact applications.

Radioterapia de salvamento para cáncer de próstata y recaída bioquímica: cinco años de experiencia. Instituto Nacional de Cancerología

Los pacientes con cáncer de próstata con tumores de riesgo bajo e intermedio de recaída pueden ser tratados con cirugía, radioterapia, y en casos seleccionados observación. Los pacientes en nuestro país, son tratados con prostatectomía radical, los cuales tienen una probabilidad de recaída bioquímica del 15% al 40% a 5 años (1,2,3). Metodología: estudio descriptivo, retrospectivo, tipo serie de casos. Se revisaron los registros de todos que recibieron radioterapia de salvamento que ofrece para a aquellos pacientes que ya tienen recaída bioquímica o local después de la Prostatectomia Radical, entre enero de 2003 y diciembre de 2007. Resultado: entre los 40 pacientes elegibles para el análisis, la media de seguimiento fue de 2,17 años, con una desviación estándar de 1,5 años, con un rango de 0 a 58 meses, la media de la edad fue de 66,12 años, con una desviación estándar de 6,63, con un rango entre 50 y 78 años. Todos los pacientes le realizaron prostatectomía. La media de supervivencia libre de enfermedad con intervalos de confianza del 95% fue de 4,58 años (2,24 a 4,92 años). Discusión: analizados los resultados en éste grupo de pacientes con cáncer de próstata sometidos a prostatectomía radical y radioterapia como terapia de salvamento, con un seguimiento promedio de 2,17 años, observamos que los resultados obtenidos en el presente estudio son inferiores a los registrados en otros reportes en la literatura (16-20).

Mutations in H5N1 influenza virus hemagglutinin that confer binding to human tracheal airway epithelium

The emergence in 2009 of a swine-origin H1N1 influenza virus as the first pandemic of the 21st Century is a timely reminder of the international public health impact of influenza viruses, even those associated with mild disease. The widespread distribution of highly pathogenic H5N1 influenza virus in the avian population has spawned concern that it may give rise to a human influenza pandemic. The mortality rate associated with occasional human infection by H5N1 virus approximates 60%, suggesting that an H5N1 pandemic would be devastating to global health and economy. To date, the H5N1 virus has not acquired the propensity to transmit efficiently between humans. The reasons behind this are unclear, especially given the high mutation rate associated with influenza virus replication. Here we used a panel of recombinant H5 hemagglutinin (HA) variants to demonstrate the potential for H5 HA to bind human airway epithelium, the predominant target tissue for influenza virus infection and spread. While parental H5 HA exhibited limited binding to human tracheal epithelium, introduction of selected mutations converted the binding profile to that of a current human influenza strain HA. Strikingly, these amino-acid changes required multiple simultaneous mutations in the genomes of naturally occurring H5 isolates. Moreover, H5 HAs bearing intermediate sequences failed to bind airway tissues and likely represent mutations that are an evolutionary "dead end." We conclude that, although genetic changes that adapt H5 to human airways can be demonstrated, they may not readily arise during natural virus replication. This genetic barrier limits the likelihood that current H5 viruses will originate a human pandemic.

Infection of human airway epithelium by human and avian strains of influenza a virus

We describe the characterization of influenza A virus infection of an established in vitro model of human pseudostratified mucociliary airway epithelium (HAE). Sialic acid receptors for both human and avian viruses, alpha-2,6- and alpha-2,3-linked sialic acids, respectively, were detected on the HAE cell surface, and their distribution accurately reflected that in human tracheobronchial tissue. Nonciliated cells present a higher proportion of alpha-2,6-linked sialic acid, while ciliated cells possess both sialic acid linkages. Although we found that human influenza viruses infected both ciliated and nonciliated cell types in the first round of infection, recent human H3N2 viruses infected a higher proportion of nonciliated cells in HAE than a 1968 pandemic-era human virus, which infected proportionally more ciliated cells. In contrast, avian influenza viruses exclusively infected ciliated cells. Although a broad-range neuraminidase abolished infection of HAE by human parainfluenza virus type 3, this treatment did not significantly affect infection by influenza viruses. All human viruses replicated efficiently in HAE, leading to accumulation of nascent virus released from the apical surface between 6 and 24 h postinfection with a low multiplicity of infection. Avian influenza A viruses also infected HAE, but spread was limited compared to that of human viruses. The nonciliated cell tropism of recent human H3N2 viruses reflects a preference for the sialic acid linkages displayed on these cell types and suggests a drift in the receptor binding phenotype of the H3 hemagglutinin protein as it evolves in humans away from its avian virus precursor.

Psychiatric disorders in children with autism spectrum disorders: prevalence, comorbidity, and associated factors in a population-derived sample

Objective: Autism spectrum disorders are now recognized to occur in up to 1% of the population and to be a major public health concern because of their early onset, lifelong persistence, and high levels of associated impairment. Little is known about the associated psychiatric disorders that may contribute to impairment. We identify the rates and type of psychiatric comorbidity associated with ASDs and explore the associations with variables identified as risk factors for child psychiatric disorders. Method: A subgroup of 112 ten- to 14-year old children from a population-derived cohort was assessed for other child psychiatric disorders (3 months' prevalence) through parent interview using the Child and Adolescent Psychiatric Assessment. DSM-IV diagnoses for childhood anxiety disorders, depressive disorders, oppositional defiant and conduct disorders, attention-deficit/hyperactivity disorder, tic disorders, trichotillomania, enuresis, and encopresis were identified. Results: Seventy percent of participants had at least one comorbid disorder and 41% had two or more. The most common diagnoses were social anxiety disorder (29.2%, 95% confidence interval [CI)] 13.2-45.1), attention-deficit/hyperactivity disorder (28.2%, 95% CI 13.3-43.0), and oppositional defiant disorder (28.1%, 95% CI 13.9-42.2). Of those with attention/deficit/hyperactivity disorder, 84% received a second comorbid diagnosis. There were few associations between putative risk factors and psychiatric disorder. Conclusions: Psychiatric disorders are common and frequently multiple in children with autism spectrum disorders. They may provide targets for intervention and should be routinely evaluated in the clinical assessment of this group.

Autistic symptomatology and language ability in autism spectrum disorder and specific language impairment

Background: Autism spectrum disorders (ASD) and specific language impairment (SLI) are common developmental disorders characterised by deficits in language and communication. The nature of the relationship between them continues to be a matter of debate. This study investigates whether the co-occurrence of ASD and language impairment is associated with differences in severity or pattern of autistic symptomatology or language profile. Methods: Participants (N = 97) were drawn from a total population cohort of 56,946 screened as part of study to ascertain the prevalence of ASD, aged 9 to 14 years. All children received an ICD-10 clinical diagnosis of ASD or No ASD. Children with nonverbal IQ 80 were divided into those with a language impairment (language score of 77 or less) and those without, creating three groups: children with ASD and a language impairment (ALI; N = 41), those with ASD and but no language impairment (ANL; N = 31) and those with language impairment but no ASD (SLI; N = 25). Results: Children with ALI did not show more current autistic symptoms than those with ANL. Children with SLI were well below the threshold for ASD. Their social adaptation was higher than the ASD groups, but still nearly 2 SD below average. In ALI the combination of ASD and language impairment was associated with weaker functional communication and more severe receptive language difficulties than those found in SLI. Receptive and expressive language were equally impaired in ALI, whereas in SLI receptive language was stronger than expressive. Conclusions: Co-occurrence of ASD and language impairment is not associated with increased current autistic symptomatology but appears to be associated with greater impairment in receptive language and functional communication.

Impairment in movement skills of children with autistic spectrum disorders

We undertook this study to explore the degree of impairment in movement skills in children with autistic spectrum disorders (ASD) and a wide IQ range. Movement skills were measured using the Movement Assessment Battery for Children (M-ABC) in a large, well defined, population-derived group of children (n=101: 89 males,12 females; mean age 11y 4mo, SD 10mo; range 10y-14y 3mo) with childhood autism and broader ASD and a wide range of IQ scores. Additionally, we tested whether a parent-completed questionnaire, the Developmental Coordination Disorder Questionnaire (DCDQ), was useful in identifying children who met criteria for movement impairments after assessment (n=97 with complete M-ABCs and DCDQs). Of the children with ASD, 79% had definite movement impairments on the M-ABC; a further 10% had borderline problems. Children with childhood autism were more impaired than children with broader ASD, and children with an IQ less than 70 were more impaired than those with IQ more than 70. This is consistent with the view that movement impairments may arise from a more severe neurological impairment that also contributes to intellectual disability and more severe autism. Movement impairment was not associated with everyday adaptive behaviour once the effect of IQ was controlled for. The DCDQ performed moderately well as a screen for possible motor difficulties. Movement impairments are common in children with ASD. Systematic assessment of movement abilities should be considered a routine investigation.

Efficacy of three screening instruments in the identification of autistic-spectrum disorders

Background Screening instruments for autistic-spectrum disorders have not been compared in the same sample. Aims To compare the Social Communication Questionnaire (SCQ), the Social Responsiveness Scale (SRS) and the Children's Communication Checklist (CCC). Method Screen and diagnostic assessments on 119 children between 9 and 13 years of age with special educational needs with and without autistic-spectrum disorders were weighted to estimate screen characteristics for a realistic target population. Results The SCQ performed best (area under receiver operating characteristic curve (AUC)=0.90; sensitivity. 6; specificity 0.78). The SRS had a lower AUC (0.77) with high sensitivity (0.78) and moderate specificity (0.67). The CCC had a high sensitivity but lower specificity (AUC=0.79; sensitivity 0.93; specificity 0.46). The AUC of the SRS and CCC was lower for children with IQ < 70. Behaviour problems reduced specificity for all three instruments. Conclusions The SCQ, SRS and CCC showed strong to moderate ability to identify autistic-spectrum disorder in this at-risk sample of school-age children with special educational needs.

Validation of the social communication questionnaire in a population cohort of children with autism spectrum disorders

Objective: To examine the properties of the Social Communication Questionnaire (SCQ) in a population cohort of children with autism spectrum disorders (ASDs) and in the general population, Method: SCQ data were collected from three samples: the Special Needs and Autism Project (SNAP) cohort of 9- to 10-year-old children with special educational needs with and without ASD and two similar but separate age groups of children from the general population (n = 411 and n = 247). Diagnostic assessments were completed on a stratified subsample (n = 255) of the special educational needs group. A sample-weighting procedure enabled us to estimate characteristics of the SCQ in the total ASD population. Diagnostic status of cases in the general population samples were extracted from child health records. Results: The SCQ showed strong discrimination between ASD and non-ASD cases (sensitivity 0.88, specificity 0.72) and between autism and nonautism cases (sensitivity 0.90, specificity 0.86). Findings were not affected by child IQ or parental education. In the general population samples between 4% and 5% of children scored above the ASD cutoff including 1.5% who scored above the autism cutoff. Although many of these high-scoring children had an ASD diagnosis, almost all (similar to 90%) of them had a diagnosed neurodevelopmental disorder. Conclusions: This study confirms the utility of the SCQ as a,first-level screen for ASD in at-risk samples of school-age children.

Prevalence of disorders of the autism spectrum in a population cohort of children in South Thames: the special needs and autism project (SNAP)

Background Recent reports have suggested that the prevalence of autism and related spectrum disorders (ASDs) is substantially higher than previously recognised. We sought to quantify prevalence of ASDs in children in South Thames, UK. Methods Within a total population cohort of 56946 children aged 9-10 years, we screened all those with a current clinical diagnosis of ASD (n=255) or those judged to be at risk for being an undetected case (n=1515). A stratified subsample (n=255) received a comprehensive diagnostic assessment, including standardised clinical observation, and parent interview assessments of autistic symptoms, language, and intelligence quotient (IQ). Clinical consensus diagnoses of childhood autism and other ASDs were derived. We used a sample weighting procedure to estimate prevalence. Findings The prevalence of childhood autism was 38.9 per 10000 (95% CI 29.9-47.8) and that of other ASDs was 77.2 per 10000 (52.1-102.3), making the total prevalence of all AS Ds 116.1 per 10000 (90.4-141.8). A narrower definition of childhood autism, which combined clinical consensus with instrument criteria for past and current presentation, provided a prevalence of 24.8 per 10 000 (17.6-32.0). The rate of previous local identification was lowest for children of less educated parents. Interpretation Prevalence of autism and related ASDs is substantially greater than previously recognised. Whether the increase is due to better ascertainment, broadening diagnostic criteria, or increased incidence is unclear. Services in health, education, and social care will need to recognise the needs of children with some form of ASD, who constitute 1% of the child population.

Measles vaccination and antibody response in autism spectrum disorders

Objective: To test the hypothesis that measles vaccination was involved in the pathogenesis of autism spectrum disorders (ASD) as evidenced by signs of a persistent measles infection or abnormally persistent immune response shown by circulating measles virus or raised antibody titres in children with ASD who had been vaccinated against measles, mumps and rubella (MMR) compared with controls. Design: Case-control study, community based. Methods: A community sample of vaccinated children aged 10-12 years in the UK with ASD (n = 98) and two control groups of similar age, one with special educational needs but no ASD (n = 52) and one typically developing group (n = 90), were tested for measles virus and antibody response to measles in the serum. Results: No difference was found between cases and controls for measles antibody response. There was no dose-response relationship between autism symptoms and antibody concentrations. Measles virus nucleic acid was amplified by reverse transcriptase-PCR in peripheral blood mononuclear cells from one patient with autism and two typically developing children. There was no evidence of a differential response to measles virus or the measles component of the MMR in children with ASD, with or without regression, and controls who had either one or two doses of MMR. Only one child from the control group had clinical symptoms of possible enterocolitis. Conclusion: No association between measles vaccination and ASD was shown.

Regression, developmental trajectory and associated problems in disorders in the autism spectrum: the SNAP study

We report rates of regression and associated findings in a population derived group of 255 children aged 9-14 years, participating in a prevalence study of autism spectrum disorders (ASD); 53 with narrowly defined autism, 105 with broader ASD and 97 with non-ASD neurodevelopmental problems, drawn from those with special educational needs within a population of 56,946 children. Language regression was reported in 30% with narrowly defined autism, 8% with broader ASD and less than 3% with developmental problems without ASD. A smaller group of children were identified who underwent a less clear setback. Regression was associated with higher rates of autistic symptoms and a deviation in developmental trajectory. Regression was not associated with epilepsy or gastrointestinal problems.