Chitin Modulates Innate Immune Responses of Keratinocytes

Chitin, after cellulose the second most abundant polysaccharide in nature, is an essential component of exoskeletons of crabs, shrimps and insects and protects these organisms from harsh conditions in their environment. Unexpectedly, chitin has been found to activate innate immune cells and to elicit murine airway inflammation. The skin represents the outer barrier of the human host defense and is in frequent contact with chitin-bearing organisms, such as house-dust mites or flies. The effects of chitin on keratinocytes, however, are poorly understood. We hypothesized that chitin stimulates keratinocytes and thereby modulates the innate immune response of the skin. Here we show that chitin is bioactive on primary and immortalized keratinocytes by triggering production of pro-inflammatory cytokines and chemokines. Chitin stimulation further induced the expression of the Toll-like receptor (TLR) TLR4 on keratinocytes at mRNA and protein level. Chitin-induced effects were mainly abrogated when TLR2 was blocked, suggesting that TLR2 senses chitin on keratinocytes. We speculate that chitin-bearing organisms modulate the innate immune response towards pathogens by upregulating secretion of cytokines and chemokines and expression of MyD88-associated TLRs, two major components of innate immunity. The clinical relevance of this mechanism remains to be defined

Dialysis-associated peritonitis in children

Peritonitis remains a frequent complication of peritoneal dialysis in children and is the most common reason for technique failure. The microbiology is characterized by a predominance of Gram-positive organisms, with fungi responsible for less than 5% of episodes. Data collected by the International Pediatric Peritonitis Registry have revealed a worldwide variation in the bacterial etiology of peritonitis, as well as in the rate of culture-negative peritonitis. Risk factors for infection include young age, the absence of prophylactic antibiotics at catheter placement, spiking of dialysis bags, and the presence of a catheter exit-site or tunnel infection. Clinical symptoms at presentation are somewhat organism specific and can be objectively assessed with a Disease Severity Score. Whereas recommendations for empiric antibiotic therapy in children have been published by the International Society of Peritoneal Dialysis, epidemiologic data and antibiotic susceptibility data suggest that it may be desirable to take the patient- and center-specific history of microorganisms and their sensitivity patterns into account when prescribing initial therapy. The vast majority of patients are treated successfully and continue peritoneal dialysis, with the poorest outcome noted in patients with peritonitis secondary to Gram-negative organisms or fungi and in those with a relapsing infection

Mutation analysis of 18 nephronophthisis associated ciliopathy disease genes using a DNA pooling and next generation sequencing strategy

Background Nephronophthisis associated ciliopathies (NPHP-AC) comprise a group of autosomal recessive cystic kidney diseases that includes nephronophthisis (NPHP), Senior-Loken syndrome (SLS), Joubert syndrome (JBTS), and Meckel-Gruber syndrome (MKS). To date, causative mutations in NPHP-AC have been described for 18 different genes, rendering mutation analysis tedious and expensive. To overcome the broad genetic locus heterogeneity, a strategy of DNA pooling with consecutive massively parallel resequencing (MPR) was devised.Methods In 120 patients with severe NPHP-AC phenotypes, five pools of genomic DNA with 24 patients each were prepared which were used as templates in order to PCR amplify all 376 exons of 18 NPHP-AC genes (NPHP1, INVS, NPHP3, NPHP4, IQCB1, CEP290, GLIS2, RPGRIP1L, NEK8, TMEM67, INPP5E, TMEM216, AHI1, ARL13B, CC2D2A, TTC21B, MKS1, and XPNPEP3). PCR products were then subjected to MPR on an Illumina Genome-Analyser and mutations were subsequently assigned to their respective mutation carrier via CEL I endonuclease based heteroduplex screening and confirmed by Sanger sequencing.Results For proof of principle, DNA from patients with known mutations was used and detection of 22 out of 24 different alleles (92% sensitivity) was demonstrated. MPR led to the molecular diagnosis in 30/120 patients (25%) and 54 pathogenic mutations (27 novel) were identified in seven different NPHP-AC genes. Additionally, in 24 patients only single heterozygous variants of unknown significance were found.Conclusions The combined approach of DNA pooling followed by MPR strongly facilitates mutation analysis in broadly heterogeneous single gene disorders. The lack of mutations in 75% of patients in this cohort indicates further extensive heterogeneity in NPHP-AC

Der Elternbildfragebogen für Kinder- und Jugendliche (EBF-KJ). Ergebnisse zur Reliabilität und Validität

Der EBF-KJ ist ein klinisch ausgerichteter Fragebogen zur Erfassung der Repräsentation der Eltern-Kind-Beziehung aus der Sicht von Kindern und Jugendlichen. Er basiert auf der systemischen Familientheorie und der Familienstresstheorie. Der Fragebogen umfasst 36 Items pro Elternteil, die sich auf drei Ressourcenskalen (Kohäsion, Identifikation, Autonomie), fünf Risikoskalen (Konflikte, Ablehnung/Gleichgültigkeit, Bestrafung, emotionale Grenzüberschreitung, Ängste/Überprotektion) und die Zusatzskala Hilfe verteilen. Der vorliegende Beitrag begründet die inhaltliche Konstruktion und stellt wichtige psychometrische Eigenschaften des EBF-KJ vor. Insgesamt wurden 785 Schüler sowie 197 Patienten einer Kinder- und Jugendpsychiatrischen Klinik im Alter von 10 bis 20 Jahren (M: 14,1) untersucht. Konfirmatorische Faktorenanalysen bestätigten die faktorielle Validität. Die interne Konsistenz lag bei 12 Skalen über .80. Weiterhin belegten systematische Korrelationen zwischen dem EBF-KJ und dem FEE (Schumacher, Eisemann & Brähler, 2000) die konvergente und diskriminante Validität beider Verfahren. Alle Skalen des EBF-KJ wiesen signifikante Zusammenhänge mit psychopathologischen Symptomen in Eltern- und Jugendfragebögen auf. ABSTRACT: Conceptually, the PRSQ is based on systemic family theory and family stress theory. The questionnaire consists of 36 items representing three resource-scales (”cohesion”, ”identification”, ”autonomy”), five risk-scales (”conflicts”, ”rejection/neglect”, ”punishment”, ”emotional burden”, ”fears/overprotection”) and one additional scale ”aid”. The study establishes the theoretical framework and presents main psychometric properties. A school-based sample of 785 participants and a clinic-referred sample of 197 outpatients aged 10 to 20 years (mean: 14.1) were surveyed. Construct validity was shown by confirmatory factor analyses. The internal consistencies of 12 of the 16 PRSQ scales were above .80. Systematic correlations between the German EMBU (FEE, Schumacher et al., 2000) and the PRSQ scales pointed to convergent and discriminant validity. Moreover, all PRSQ scales turned out to be significantly correlated with psychopathological symptoms as measured by questionnaires

Desperately seeking diarrhoea: outbreak of haemolytic uraemic syndrome caused by emerging sorbitol-fermenting shiga toxin-producing Escherichia coli O157:H-, Germany, 2009

Infection with sorbitol-fermenting Shiga toxin-producing Escherichia coli O157:H- (sf STEC O157:H-) is rare, but emerging in Europe. The pathogen is typically isolated from paediatric patients with life-threatening haemolytic uraemic syndrome (HUS). It is unclear whether this observation primarily reflects the pathogen's virulence or its complex laboratory diagnosis, not routinely conducted in diarrhoeal patients. In summer 2009, four boys living in the same suburb in Germany developed diarrhoea-associated HUS: three were infected by sf STEC O157:H- and one died. We conducted two analytical epidemiological studies, an extensive search for diarrhoeal cases in potentially exposed groups, and an environmental investigation. Outbreak cases were residents of the suburb diagnosed with HUS, sf STEC O157:H- infection, or both between 24 July 2009 and 25 August 2009. Overall, we ascertained eight cases with a median age of 4 years (range: from 8 months to 9 years). Stool screening of 220 persons led to the identification of only four additional cases: two asymptomatic carriers and two diarrhoeal cases. HUS was strongly associated with visiting a local playground in July, particularly on 16th July (odds ratio = 42.7, P = 0.002). No other commonality, including food, was identified, and all environmental samples (n = 24) were negative. In this localized non-foodborne outbreak, the place of likely infection was a local playground. Sf STEC O157:H- infection apparently limits itself rarely to diarrhoeal illness and progresses frequently to HUS. Therefore, detection of and response to this hypervirulent pathogen primarily relies on HUS surveillance

Shiga Toxin-Producing Escherichia coli Infection and Antibodies against Stx2 and Stx1 in Household Contacts of Children with Enteropathic Hemolytic-Uremic Syndrome

Ninety-five household contacts (aged 2 months to 73 years) of patients with enteropathic hemolytic-uremic syndrome (HUS) were investigated for the presence of immunoglobulin (Ig) G antibodies to Shiga toxins Stx2 and Stx1 by Western blot assay. Thirty-one percent of the household contacts and 19% of 327 controls had anti-Stx2 IgG (heavy and light chain [H + L]), 5 and 8%, respectively, had anti-Stx1 IgG (H + L), and 3 and 2%, respectively, had both anti-Stx2 and anti-Stx1 IgG (H + L). The incidence of infections with Stx-producing Escherichia coli (STEC) was determined based on the following diagnostic criteria: STEC isolation, detection of stx gene sequences, free fecal Stx in stool filtrates, and serum IgM antibodies against E. coli O157 lipopolysaccharide. Evidence of STEC infection was observed in 25 household contacts, of whom 18 (72%) were asymptomatic and represented a potential source of infection. Six of 13 (46%) household contacts with Stx2-producing E. coli O157:H7 in stool culture developed anti-Stx2 IgG (H + L), compared to 71% of Stx2-associated HUS cases. In individuals showing anti-Stx2 IgG (H + L), the antibody response was directed against the B subunit in 69% of household contacts and 71% of controls, in contrast to 28% of HUS patients. In this investigation controls had a significant increase of the median of IgM antibodies to O157 lipopolysaccharide (LPS) with age, up to the fifth decade. The lack of disease in household contacts with B subunit-specific antibodies, as well as the significantly higher median of anti-O157 LPS IgM antibodies in controls beyond 4.9 years of age, suggests a protective role for anti-Stx and anti-O157 LPS antibodies

The command to love the neighbour in Paul and the Synoptics

G.J.S. is the supervisor of H.A.H.’s PhD thesis called ‘Paul’s use of the Synoptic Jesus tradition’.When sayings of Jesus are compared between the Pauline letters and the Synoptic Gospels in an attempt to locate parallels, Galatians 5:14 and Romans 13:8–10 have frequently been put forward as possible parallels to the Synoptic renditions of the great commandment of Mark 12:28–34 (Mt 22:34–40; Lk 10:25–28). These Pauline and Synoptic texts all contain the command to love the neighbour, but the Synoptic texts have added the command to love God to the command to love the neighbour. Paul never quoted the great commandment. Consequently, a relationship between the verses is normally rejected. However, not all possibilities have been explored. In the search for parallels between Pauline and Synoptic Jesus traditions it has been overlooked that Paul and Matthew render the command to love the neighbour more than once. Matthew delivers the command to love the neighbour three times. Only once he has connected it to the command to love God. Matthew renders the single command to love the neighbour twice, resembling the Pauline use of the command. Using the criteria for validating parallels between Pauline and Synoptic Jesus traditions, including verbal agreement and the unique use of the command to love the neighbour by Paul and Matthew, it is argued that a connection between Romans 13:9 and Matthew 19:18–19 is likely. INTRADISCIPLINARY AND/OR INTERDISCIPLINARY IMPLICATIONS : The research explores the use of Leviticus 19:18b by the New Testament authors. It is argued that the command to love the neighbour was given high prominence in the early church, as it was used by the Synoptic authors and by Paul to summarise Jesus’ ethical teachings.http://www.ve.org.zaam2016New Testament Studie