High producer variant of lipoprotein lipase may protect from hepatocellular carcinoma in alcohol-associated cirrhosis.

Background & Aims: Progression of alcohol-associated liver disease (ALD) is driven by genetic predisposition. The rs13702 variant in the lipoprotein lipase (LPL) gene is linked to non-alcoholic fatty liver disease. We aimed at clarifying its role in ALD. Methods: Patients with alcohol-associated cirrhosis, with (n = 385) and without hepatocellular carcinoma (HCC) (n = 656), with HCC attributable to viral hepatitis C (n = 280), controls with alcohol abuse without liver damage (n = 366), and healthy controls (n = 277) were genotyped regarding the LPL rs13702 polymorphism. Furthermore, the UK Biobank cohort was analysed. LPL expression was investigated in human liver specimens and in liver cell lines. Results: Frequency of the LPL rs13702 CC genotype was lower in ALD with HCC in comparison to ALD without HCC both in the initial (3.9% vs. 9.3%) and the validation cohort (4.7% vs. 9.5%; p <0.05 each) and compared with patients with viral HCC (11.4%), alcohol misuse without cirrhosis (8.7%), or healthy controls (9.0%). This protective effect (odds ratio [OR] = 0.5) was confirmed in multivariate analysis including age (OR = 1.1/year), male sex (OR = 3.0), diabetes (OR = 1.8), and carriage of the PNPLA3 I148M risk variant (OR = 2.0). In the UK Biobank cohort, the LPL rs13702 C allele was replicated as a risk factor for HCC. Liver expression of LPL mRNA was dependent on LPL rs13702 genotype and significantly higher in patients with ALD cirrhosis compared with controls and alcohol-associated HCC. Although hepatocyte cell lines showed negligible LPL protein expression, hepatic stellate cells and liver sinusoidal endothelial cells expressed LPL. Conclusions: LPL is upregulated in the liver of patients with alcohol-associated cirrhosis. The LPL rs13702 high producer variant confers protection against HCC in ALD, which might help to stratify people for HCC risk. Impact and implications: Hepatocellular carcinoma is a severe complication of liver cirrhosis influenced by genetic predisposition. We found that a genetic variant in the gene encoding lipoprotein lipase reduces the risk for hepatocellular carcinoma in alcohol-associated cirrhosis. This genetic variation may directly affect the liver, because, unlike in healthy adult liver, lipoprotein lipase is produced from liver cells in alcohol-associated cirrhosis.

Genetic Variation of SAMM50 Is Not an Independent Risk Factor for Alcoholic Hepatocellular Carcinoma in Caucasian Patients.

Hepatocellular carcinoma (HCC) is a severe complication of advanced alcoholic liver disease, which is modulated by genetic predisposition. Identifying new genetic loci might improve screening. Genetic variation of SAMM50 was linked to HCC. We aimed to validate this finding in a large cohort of patients with advanced alcoholic liver disease (ALD). A large, well-characterised cohort of patients with alcoholic cirrhosis without (n = 674) and with (n = 386) HCC, as well as controls with HCC due to viral hepatitis (n = 134), controls with heavy alcohol abuse without liver disease (n = 266) and healthy subjects (n = 237), were genotyped for SAMM50 rs3827385 and rs3761472 and for PNPLA3 rs738409. Genotype frequencies were compared between patients with alcohol-associated cirrhosis with and without HCC by uni- and multivariate analysis. Minor variants in both SAMM50 rs3827385 and rs3761472 were significantly more frequent in patients with alcoholic HCC versus alcoholic cirrhosis and versus the control cohorts. An even stronger association was noted for PNPLA3 rs738409. The univariate analysis resulted in an odds ratio (OR) of 1.8 for carriers of at least one minor variant of SAMM50 rs3827385 and rs3761472 (each p < 0.001), but this association was lost in multivariate analysis with age (OR 1.1/year), male sex (OR 3.2), diabetes (OR 1.9) and carriage of PNPLA3 148M (OR 2.1) remaining in the final model. Although minor variants of both SAMM50 loci are strongly associated with alcoholic HCC, this association is not independent of carriage of the well-known risk variant PNPLA3 148M.

Facilitators and Barriers for Young Medical Doctors Writing Their First Manuscript for Publication.

Although scientific publication is often mandatory in medical professions, writing the first research article for publication is challenging, especially as medical curricula have only a minor focus on scientific writing. The aim was therefore to identify facilitators and barriers experienced by medical doctors writing their first scientific article for publication. An explorative inductive approach made use of semi-structured interviews for collecting data until saturation. Data were analyzed with systematic text condensation. Several barriers were identified: (a) writing in general; (b) writing in English; (c) dealing with content, structure, and presentation; and (d) navigating in the author group. Good supervision in the initial writing phase was a facilitating factor. Medical doctors requested a course in which they could work on their own articles and give feedback to fellow students. They valued skilled lecturers and individual supervision, and they wanted to learn about author instructions, how to present text correctly, and how to sell their core message. Their goal was to create a useful end product and to obtain European Credit Transfer System (ECTS) points. The facilitators and barriers that medical doctors experience when writing their first scientific article for publication and their course requests should be reflected in the learning objectives and content of future courses.

Von Willebrand factor, its cleaving protease (ADAMTS13), and inflammation in young adults: The African-PREDICT study.

BACKGROUND: The role of inflammation in the early development of vascular dysfunction remains complex. Interleukin-6 (IL-6) and C-reactive protein (CRP) can cause an acute imbalance in the von Willebrand factor (vWF)-ADAMTS13 interaction, indicating a possible link between markers of haemostasis and low-grade inflammation. To better understand these inter-relationships in the early phases of disease development, we investigated whether vWF and ADAMTS13 associate with the pro-inflammatory markers, IL-6 and CRP in healthy young adults. We considered the role of blood types, sex and race on these relationships. METHODS: In healthy black and white men and women (n = 1113; 24 ± 5 years; no previous diagnosis or medication use for chronic diseases) we analysed von Willebrand factor antigen (vWFag), ADAMTS13, IL-6 and CRP, and grouped blood types as non-O (A, B and AB) and O. Covariates included socioeconomic status, age, estimated glomerular filtration rate, 24-hour systolic blood pressure, waist circumference, glucose, total cholesterol, platelet count, γ-glutamyl transferase and total energy expenditure. RESULTS: In the total group, vWFag was highest in the third tertile of both IL-6 and CRP (p ≤ 0.014), while ADAMTS13 was lowest in the third compared to the first IL-6 tertile (p = 0.006). In multivariate regression, vWFag associated positively with IL-6 (Adj R2 = 0.169; ß = 0.123; p = 0.001) and CRP (Adj R2 = 0.163; ß=0.094; p = 0.019) in the total group, in the O blood group (all p ≤ 0.051) and white men (all p ≤ 0.035). ADAMTS13 associated negatively with IL-6 (Adj R2 = 0.053; ß = -0.154; p = 0.015) and CRP (Adj R2 = 0.055; ß = -0.177; p = 0.009), only in the O blood group. CONCLUSIONS: Markers of haemostasis associated independently with low-grade inflammation in the O type blood group and white men. An interplay between the haemostatic and inflammatory systems may already exist in young healthy adults and is dependent on blood groups, sex and race. This extends our understanding on the role of inflammation in the early development of vascular dysfunction prior to cardiovascular compromise.

[Demographic characteristics of foster families in Germany]. / Demografische Eigenschaften von Pflegefamilien in Deutschland.

Demographic characteristics of foster families in Germany Abstract. Objective: Demographic characteristics like the level of education, job position or the distribution of age in families have a significant impact on the development of children. Therefore, in the current study, we examined the commonalities and differences of foster family samples, recruited for research, with the general population. Method: The data at hand are part of the "GROW&TREAT" project that examines the development of foster children in comparison to children who live with their biological families. Results: Differences were found in comparison to the data of the German Census Bureau for the foster family sample and the biological sample for parental education, apportionment of parental employment, and familial net income. Concerning the direct sample group comparison, differences could be observed in terms of age gap between (foster) mother and child and parental education. Furthermore, the representativeness of the foster family sample was analyzed based on reports of participating youth welfare services. Conclusions: The examined sample seems to be representative for foster family samples in research contexts. The consequences of these differences on potential research results and recruitment characteristics were further discussed.

Children's mental health problems and their relation to parental stress in foster mothers and fathers.

BACKGROUND: This study focuses on children living in foster families with a history of maltreatment or neglect. These children often show adverse mental health outcomes reflected in increased externalizing and internalizing problems. It is expected that these adverse outcomes are associated with increased parental stress levels experienced by foster mothers as well as foster fathers. METHODS: The study sample included 79 children living in foster families and 140 children living in biological families as comparison group. The age of the children ranged from 2 to 7 years. Mental health problems were assessed with the Child Behavior Checklist, while parenting stress was measured with a parenting stress questionnaire including subscales on the amount of experienced stress and the amount of perceived support. The Child Behavior Checklist assessments were based mainly on maternal reports, while the parental stress assessments were based on maternal as well as paternal reports. RESULTS: As expected the results showed increased externalizing and internalizing scores for the foster children accompanied by increased parental stress experiences in the foster family sample (however only in the maternal, but not in the paternal stress reports). The stress differences between the foster and biological family groups disappeared, when the children's mental health problem scores were included as covariates. Moreover, especially the externalizing scores were strong predictors of parental stress in both, the groups of foster and biological parents. The amount of perceived social support was associated with reduced parental stress, but only in the group of biological fathers. CONCLUSION: The emergence of parental stress in biological as well as foster parents is closely related to child characteristics (mainly externalizing child problems). Possible implications for the reduction of parental stress are discussed as a consequence of the present results.

Bcl-2 and Bcl-XL are indispensable for the late phase of mast cell development from mouse embryonic stem cells.

OBJECTIVE: The aim of this study was to determine the importance of the prosurvival factors Bcl-2 and Bcl-XL for mast cell development and survival. METHODS: bcl-x(-/-) and bcl-2(-/-) mouse embryonic stem cells were maintained in medium supplemented with either interleukin (IL)-3 or IL-3 in combination with stem cell factor (SCF) to favor mast cell development. The development of Bcl-2 family deficient embryonic stem cell-derived mast cells (ESMCs) was monitored and Bcl-2 family gene expression and cell numbers were analyzed. RESULTS: Deficiency in either bcl-x or bcl-2 totally inhibited the development of ESMCs when IL-3 alone was used as a mast cell growth factor. Intriguingly, when IL-3 was used in combination with SCF, the ESMCs developed normally the first 2 weeks but thereafter the cell numbers dropped drastically. The remaining ESMCs express mouse mast cell protease 1, suggesting a mucosal-like phenotype. ESMCs lacking bcl-x or bcl-2 exhibited strong expression of A1, another prosurvival Bcl-2 family member. CONCLUSION: For the first time we provide direct evidence that both bcl-x and bcl-2 are indispensable for mast cell survival during the late phase of their development.

Mast cell CD30 ligand is upregulated in cutaneous inflammation and mediates degranulation-independent chemokine secretion.

Mast cells are involved in many disorders where the triggering mechanism that leads to degranulation and/or cytokine secretion has not been defined. Several chronic inflammatory diseases are associated with increased mast cell numbers and upregulation of the TNF receptor family member CD30, but the role of elevated CD30 expression is poorly understood. Here we report what we believe to be a novel way to activate mast cells with CD30 that leads to degranulation-independent secretion of chemokines. CD30 induced a de novo synthesis and secretion of the chemokines IL-8, macrophage inflammatory protein-1alpha (MIP-1alpha), and MIP-1beta, a process involving the MAPK/ERK pathway. Mast cells were found to be the predominant CD30 ligand-positive (CD30L-positive) cell in the chronic inflammatory skin diseases psoriasis and atopic dermatitis, and both CD30 and CD30L expression were upregulated in lesional skin in these conditions. Furthermore, the number of IL-8-positive mast cells was elevated both in psoriatic and atopic dermatitis lesional skin as well as in ex vivo CD30-treated healthy skin organ cultures. In summary, characterization of CD30 activation of mast cells has uncovered an IgE-independent pathway that is of importance in understanding the entirety of the role of mast cells in diseases associated with mast cells and CD30 expression. These diseases include Hodgkin lymphoma, atopic dermatitis, and psoriasis.

Readministration of IgE is required for repeated passive cutaneous anaphylaxis in mice.

BACKGROUND: Homologous passive cutaneous anaphylaxis (PCA) is an established technique that provides a useful tool to study local inflammation due to mast cell activation. Since mast cells are responsive to repeated challenge by IgE receptor crosslinking in vitro, we investigated the responsiveness of mice to repeated PCA activation. METHODS: Female BALB/c mice were sensitized with intradermal injection of IgE into the ear and provocated through intravenous injection of trinitrophenol-BSA. Repeated provocation was administered, and the outcome was determined by colorimetrical measurement of Evans blue dye leakage. RESULTS: After local IgE injection, mice could be challenged to demonstrate a PCA reaction at least up to 13 days after IgE sensitization. In contrast, passively sensitized mice did not respond to repeated antigen challenge, i.e. a second provocation administered between 1 and 12 days after the first PCA reaction. However, if these mice were intradermally resensitized with IgE at the same site after the first challenge, they became responsive to the repeated challenge. CONCLUSIONS: Mast cells become desensitized upon PCA reaction, and resensitization with IgE is critical for mice repeatedly stimulated by PCA activation.

Coaggregation of FcepsilonRI with FcgammaRIIB Inhibits Degranulation but Not Induction of Bcl-2 Family Members A1 and Bim in Mast Cells.

: The aggregation of high-affinity immunoglobulin E (IgE) receptors (FcepsilonRI) on mast cells is a critical event in the initiation of an allergic reaction. Coengagement of FcepsilonRI with immunoglobulin G (IgG) low-affinity receptor FcgammaRIIB/CD32 inhibits degranulation and the release of inflammatory mediators from mast cells and has therefore been proposed as a new therapeutic approach for the treatment of allergies. In this study, we investigated whether FcgammaRIIB, besides inhibiting degranulation, negatively regulates other signalling pathways downstream of FcepsilonRI. For this, we determined the phosphorylation and/or expression of proteins involved in the regulation of mast-cell apoptosis. Coaggregation led to an attenuation of Akt phosphorylation but did not inhibit phosphorylation of transcription factor Foxo3a or its proapoptotic target, Bim. Similarly, FcepsilonRI-dependent expression of the prosurvival gene A1 was not affected by coaggregation. Our data demonstrate that coengagement of FcepsilonRI and FcgammaRIIB inhibits degranulation but not the signalling pathways regulating Bcl-2 family members Bim and A1.

Stem cell factor promotes mast cell survival via inactivation of FOXO3a-mediated transcriptional induction and MEK-regulated phosphorylation of the proapoptotic protein Bim.

Mast cells are found in tissues throughout the body where they play important roles in the regulation of inflammatory responses. One characteristic feature of mast cells is their longevity. Although it is well established that mast cell survival is dependent on stem cell factor (SCF), it has not been described how this process is regulated. Herein, we report that SCF promotes mast cell survival through inactivation of the Forkhead transcription factor FOXO3a (forkhead box, class O3A) and down-regulation and phosphorylation of its target Bim (Bcl-2 [B-cell lymphoma-2] interacting modulator of cell death), a Bcl-2 homology 3 (BH3)-only proapoptotic protein. SCF induced a rapid and transient phosphorylation of Akt (protein kinase B) and FOXO3a. SCF treatment prevented up-regulation of Bim protein expression and led to increased Bim phosphorylation. Bim phosphorylation was inhibited by PD98059 and LY294002 treatment, suggesting the involvement of mitogen-activated protein kinase kinase/mitogen-activated protein kinase (MEK/MAPK) and phosphatidylinositol 3 (PI3)-kinase pathways in this process. Overexpression of phosphorylation-deficient FOXO3a caused an up-regulation of Bim and induced mast cell apoptosis even in the presence of SCF. Mast cell apoptosis induced by the phosphorylation-deficient FOXO3a was attenuated in bim-/- mast cells. Because apoptosis is abnormally reduced in bim-/- mast cells, these data provide evidence that Akt-mediated inhibition of FOXO3a and its transcription target Bim provides an important mechanism by which SCF acts to prevent apoptosis in mast cells.