Molekulargenetische und klinische Untersuchungen zur Laktoseintoleranz (Hypolaktasie)

2002 wurde mit dem Genotypen LCT-13910C/C ein Marker beschrieben, der mit dem primären adulten Laktasemangel (PAL) assoziiert sein soll. Ziel war die Evaluierung des entsprechenden genetischen Tests. Referenz war der H2-Atemtest. Die Studie bestätigte mit einer Sensitivität und Spezifität von 89% bzw. 97% für den genetischen Test, dass sich das Verfahren im klinischen Einsatz zum Ausschluss eines PAL eignet. Zudem erfolgte in einer Genotypisierungsstudie die Erhebung der Prävalenz des PAL erstmalig im Raum Rostock und zu interethnischen Vergleichszwecken in einer asiatischen Kohorte

Neurodevelopmental problems in maltreated children referred with indiscriminate friendliness

We aimed to explore the extent of neurodevelopmental difficulties in severely maltreated adopted children. We recruited 34 adopted children, referred with symptoms of indiscriminate friendliness and a history of severe maltreatment in their early childhood and 32 typically developing comparison children without such a history, living in biological families. All 66 children, aged 5–12 years, underwent a detailed neuropsychiatric assessment. The overwhelming majority of the adopted/indiscriminately friendly group had a range of psychiatric diagnoses, including Attention Deficit Hyperactivity Disorder (ADHD), Post-Traumatic Stress Disorder (PTSD) and Reactive Attachment Disorder (RAD) and one third exhibited the disorganised pattern of attachment. The mean IQ was 15 points lower than the comparison group and the majority of the adopted group had suspected language disorder and/or delay. Our findings show that school-aged adopted children with a history of severe maltreatment can have very complex and sometimes disabling neuropsychiatric prob

An exploratory study of the association between reactive attachment disorder and attachment narratives in early school-age children

To explore attachment narratives in children diagnosed with reactive attachment disorder (RAD). Method: We compared attachment narratives, as measured by the Manchester Child Attachment Story Task, in a group of 33 children with a diagnosis of RAD and 37 comparison children. Results: The relative risk (RR) for children with RAD having an insecure attachment pattern was 2.4 (1.4-4.2) but 30% were rated as securely attached. Within the RAD group, children with a clear history of maltreatment were more likely to be Insecure-Disorganised than children without a clear history of maltreatment. Conclusions: Reactive attachment disorder is not the same as attachment insecurity, and questions remain about how attachment research informs clinical research on attachment disorders

Children’s emotion understanding in relation to attachment to mother and father

This is the author accepted manuscript. The final version is available from Wiley via the DOI in this record.Although attachment plays a key role in children’s socioemotional development, little attention has been paid to the role of children’s attachment to their father. This study examined whether insecure attachment to each parent was associated with reduced emotion understanding in children and whether children showed consistent attachments to their mother and father. We measured children’s attachment to each parent using the Manchester Child Attachment Story Task and child emotion understanding using the Test of Emotion Comprehension (children’s Mage = 5.64 years, SD = 0.84). The results indicated that insecure father-child attachment and insecure mother-child attachment were each associated with lower emotion understanding in children after controlling for parent’s depressive symptoms and children’s age. There was significant concordance of child attachment to mother and father. The findings provide support for convergence of children’s attachment across parents and suggest that father-child attachment is an important factor to consider when examining children’s emotion understanding.We would like to thank all families who took part in this research. The study was funded by a British Academy small grant awarded to LP and WK (SG112244

High sensitive troponin T and heart fatty acid binding protein: Novel biomarker in heart failure with normal ejection fraction?: A cross-sectional study

Background: High sensitive troponin T (hsTnT) and heart fatty acid binding protein (hFABP) are both markers of myocardial injury and predict adverse outcome in patients with systolic heart failure (SHF). We tested whether hsTnT and hFABP plasma levels are elevated in patients with heart failure with normal ejection fraction (HFnEF). Methods: We analyzed hsTnT, hFABP and N-terminal brain natriuretic peptide in 130 patients comprising 49 HFnEF patients, 51 patients with asymptomatic left ventricular diastolic dysfunction (LVDD), and 30 controls with normal diastolic function. Patients were classified to have HFnEF when the diagnostic criteria as recommended by the European Society of Cardiology were met. Results: Levels of hs TnT and hFABP were significantly higher in patients with asymptomatic LVDD and HFnEF (both p < 0.001) compared to controls. The hsTnT levels were 5.6 [0.0-9.8] pg/ml in LVDD vs. 8.5 [3.9-17.5] pg/ml in HFnEF vs. < 0.03 [< 0.03-6.4] pg/ml in controls; hFABP levels were 3029 [2533-3761] pg/ml in LVDD vs. 3669 [2918-4839] pg/ml in HFnEF vs. 2361 [1860-3081] pg/ml in controls. Furthermore, hsTnT and hFABP levels were higher in subjects with HFnEF compared to LVDD (p = 0.015 and p = 0.022). Conclusion: In HFnEF patients, hsTnT and hFABP are elevated independent of coronary artery disease, suggesting that ongoing myocardial damage plays a critical role in the pathophysiology. A combination of biomarkers and echocardiographic parameters might improve diagnostic accuracy and risk stratification of patients with HFnEF

ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing

Despite intense interest, methods that provide enhanced sensitivity and specificity in parallel measurements of candidate protein biomarkers in numerous samples have been lacking. We present herein a multiplex proximity ligation assay with readout via realtime PCR or DNA sequencing (ProteinSeq). We demonstrate improved sensitivity over conventional sandwich assays for simultaneous analysis of sets of 35 proteins in 5 µl of blood plasma. Importantly, we observe a minimal tendency to increased background with multiplexing, compared to a sandwich assay, suggesting that higher levels of multiplexing are possible. We used ProteinSeq to analyze proteins in plasma samples from cardiovascular disease (CVD) patient cohorts and matched controls. Three proteins, namely P-selectin, Cystatin-B and Kallikrein-6, were identified as putative diagnostic biomarkers for CVD. The latter two have not been previously reported in the literature and their potential roles must be validated in larger patient cohorts. We conclude that ProteinSeq is promising for screening large numbers of proteins and samples while the technology can provide a much-needed platform for validation of diagnostic markers in biobank samples and in clinical use

Development of a method by gas chromatography and triple quadrupole mass spectrometry for the quantification of estrogen metabolites in human breast tissue

Im Rahmen der Arbeit wurde eine Methode für die Quantifizierung von freiem 17β-Estradiol, Estron sowie der hydroxylierten und methylierten Metabolite im Brustgewebe entwickelt. Aufgrund der geringen Probengehalte erforderte dies eine gezielte Isolierung der Analyte aus der Probenmatrix sowie eine effektive Aufreinigung und Aufkonzentrierung, so dass eine Extraktion mit anschließender Festphasenextraktion durchgeführt wurde. Zudem wurde eine empfindliche Mess-Methode etabliert, welche auf Grundlage einer multi-reaction-monitoring-Methode, mittels Gaschromatographie und gekoppelten Triple-Quadrupol-Massenspektrometer, entwickelt wurde. Die Anwendbarkeit der Aufarbeitungs- und Mess-Methode wurde überprüft, indem diese auf 30 Realproben übertragen wurde. Dabei sind die ermittelten Gehalte mit den publizierten Daten der Gewebekonzentrationen von 17β-Estradiol, Estron und deren Metaboliten verglichen und Korrelationen mit ausgewählten Brustkrebs-begünstigenden Risikofaktoren betrachtet worden. Um ein quantitatives Metabolitenprofil von 17β-Estradiol, Estron und deren Metaboliten im Gewebe zu erstellen, wurden mit Hilfe einer multi-reaction-monitoring-Methode für alle Metabolite ein spezifischer Quanti- und Qualifier-Übergang etabliert. Durch die Optimierung der Ionisierungs- und Kollisionsenergien sowie der Initial-, Transferline- und Ionenquell-Temperatur beziehungsweise der dwell-time wurden Methoden- und Geräte-bedingte Empfindlichkeitsverluste so weit wie möglich reduziert, so dass maximale Signalintensitäten aller Quantifier-Übergänge gewährleistet waren. Zur gezielten Isolation sowie Aufreinigung und Anreicherung der Analyten,... ...so dass trotz der geringen Anzahl analysierter Gewebe-spenden der Einfluss des Body-Mass-Index und die Einnahme oraler Kontrazeptiva auf die Gehalte von 17β-Estradiol in der prämenopausalen Frau deutlich wurden. Die entwickelte Mess-Methode ermöglicht den routinemäßigen Einsatz für die Quantifizierung von freiem 17β-Estradiol, Estron und deren Methyl-Catecholen in humanem Brustgewebe. Beim Vergleich der berechneten Nachweisgrenzen von Catechol-Estrogenen mit Literaturangaben wurde herausgestellt, dass empfindlichere flüssigchromatographische Methoden als Methode der Wahl bei deren Analytik heranzuziehen sind. Die Übertragung der in Standardlösungen durchgeführten Versuche zur enzymatischen Hydrolyse von Glucuronid-und Sulfat-Konjugaten auf Gewebematrix stellt für weiterführende Arbeiten den entscheidenden Ansatzpunkt dar, um ein quantitatives Metabolitenprofil von freiem und gebundenem 17β-Estradiol, Estron und den Metaboliten in Brustgewebe erstellen zu können.The aim of the present work was to develop a method for the quantification of free estradiol, estrone, catechol estrogens and methylation products in breast tissue. Due to trace amounts in the samples it was necessary to isolate the analytes from the sample matrix, purify and concentrate them prior to tissue extraction and solid phase extraction. Additionally, a sensitive method of measurement based on multi reaction monitoring using gas chromatography and triple quadrupole mass spectrometry was developed. The applicability of the sample preparation and measurement method was tested in 30 tissue samples by comparing the measured levels with published data concerning concentrations of estradiol, estrone and their metabolites in breast tissue. Also it was necessary to find possible correlations between the analytes concentrations and selected risk factors for breast cancer. In order to develop a quantitative profile of estradiol, estrone and metabolites in tissue, qualifier and quantifier transitions were established using the multi reaction monitoring mode. By optimising the ionisation and collision energy, the initial, transferline and ion source temperatures and the dwell-time, it was possible to reduce the loss of sensitivity caused by both method and equipment and maximize the signal intensity of all quantifier transitions. For the isolation, purification and concentration of the analytes... ...have some influence on the levels of estradiol in premenopausal women. The developed measurement method enabled a routine usage for the quantification of free estradiol, estrone and methylcatechols in human breast tissue. The comparison between the calculated detection levels of catechol estrogens and the published data led to the conclusion that liquid chromatographic methods are more sensitive. The transference of these methods and experiments conducted, such as for the enzymatic hydrolysis of glucuronide- and sulfate-conjugates on tissue matrix in standard solution, seem to offer a starting point for future research projects. Especially, for the purpose of establishing a quantitative metabolite profile of free and conjugated estradiol, estrone and their metabolites

Obsessive-compulsive disorder in children and adolescents: Parental understanding, accommodation, coping and distress

Parental accommodation of pediatric OCD is common and is associated with negative affect in parents. Qualitative accounts of caring for a child with OCD are limited and no studies have assessed differences between mothers and fathers in accommodation, coping and distress. The current study used a mixed methods approach to understand parental accommodation, negative affect and coping. Forty-one mothers and 29 fathers of 43 children with OCD were asked to write narratives about their understanding and management of OCD and to complete measures of accommodation, coping, and distress. Symptom accommodation was high with almost half of the parents watching the child complete rituals or waiting for the child on a daily basis. Analysis of parental narratives indicated a distressing struggle between engaging in and resisting accommodation in order to manage their own and their child’s anger and distress. T-tests and correlation analysis indicated that accommodation did not differ significantly between mothers and fathers but was more strongly associated with negative affect in mothers. Analyses indicated that mothers reported using all types of coping strategy more often than fathers, particularly escape-avoidance, taking responsibility and using social support. Escape-avoidance coping was positively correlated with accommodation and negative affect in both mothers and fathers. Interventions that target parental constructions of OCD and their behavioural and emotional responses to it may assist in reducing the occurrence of accommodation, avoidant coping and parental distress

Obsessive-compulsive disorder in children and adolescents: parental understanding, accommodation, coping and distress

Parental accommodation of pediatric OCD is common and is associated with negative affect in parents. Qualitative accounts of caring for a child with OCD are limited and no studies have assessed differences between mothers and fathers in accommodation, coping and distress. The current study used a mixed methods approach to understand parental accommodation, negative affect and coping. Forty-one mothers and 29 fathers of 43 children with OCD were asked to write narratives about their understanding and management of OCD and to complete measures of accommodation, coping, and distress. Symptom accommodation was high with almost half of the parents watching the child complete rituals or waiting for the child on a daily basis. Analysis of parental narratives indicated a distressing struggle between engaging in and resisting accommodation in order to manage their own and their child's anger and distress. T-tests and correlation analysis indicated that accommodation did not differ significantly between mothers and fathers but was more strongly associated with negative affect in mothers. Analyses indicated that mothers reported using all types of coping strategy more often than fathers, particularly escape-avoidance, taking responsibility and using social support. Escape-avoidance coping was positively correlated with accommodation and negative affect in both mothers and fathers. Interventions that target parental constructions of OCD and their behavioural and emotional responses to it may assist in reducing the occurrence of accommodation, avoidant coping and parental distress