Bernhard Vaske

Prospective biopsy-controlled evaluation of cell death biomarkers for prediction of liver fibrosis and nonalcoholic steatohepatitis

Fibrosis and steatosis are major histopathological alterations in chronic liver diseases. Despite various shortcomings, disease severity is generally determined by liver biopsy, emphasizing the need for simple noninvasive methods for assessing disease activity. Because hepatocyte cell death is considered a crucial pathogenic factor, we prospectively evaluated the utility of serum biomarkers of cell death to predict different stages of fibrosis and steatosis in 121 patients with chronic liver disease. We compared the M30 enzyme‐linked immunosorbent assay (ELISA), which detects a caspase‐cleaved cytokeratin‐18 (CK‐18) fragment and thereby apoptotic cell death, with the M65 ELISA, which detects both caspase‐cleaved and uncleaved CK‐18 and thereby overall cell death. Both biomarkers significantly discriminated patients with different fibrosis stages from healthy controls. However, whereas both markers differentiated low or moderate from advanced fibrosis, only the M65 antigen could discriminate even lower stages of fibrosis. The M65 assay also performed better in distinguishing low (≤10%) and higher (>10%) grades of steatosis. In a subgroup of patients, we evaluated the biomarkers for their power to predict nonalcoholic steatohepatitis (NASH). Importantly, both markers accurately differentiated healthy controls or simple steatosis from NASH. However, only serum levels of M65 antigen could differentiate simple steatosis from healthy controls. Conclusion: Cell death biomarkers are potentially useful to predict fibrosis, steatosis, or NASH. Compared with the widely used apoptosis marker M30, the M65 assay had a better diagnostic performance and even differentiated between lower fibrosis stages as well as between healthy individuals and patients with simple steatosis. (HEPATOLOGY 2012)

IL-6 predicts organ dysfunction and mortality in patients with multiple injuries

BackgroundAlthough therapeutic concepts of patients with major trauma have improved during recent years, organ dysfunction still remains a frequent complication during clinical course in intensive care units. It has previously been shown that cytokines are upregulated under stress conditions such as trauma or sepsis. However, it is still debatable if cytokines are adequate parameters to describe the current state of trauma patients. To elucidate the relevance of cytokines, we investigated if cytokines predict development of multiple organ dysfunction syndrome (MODS) or outcome.MethodsA total of 143 patients with an injury severity score ≥ 16, between 16 and 65 years, admitted to the Hannover Medical School Level 1 Trauma Center between January 1997 and December 2001 were prospectively included in this study. Marshall Score for MODS was calculated for at least 14 days and plasma levels of TNF-α, IL-1β, IL-6, IL-8 and IL-10 were measured. To determine the association between cytokine levels and development of MODS the Spearman rank correlation coefficient was calculated and logistic regression and analysis were performed.Results and DiscussionPatients with MODS had increased plasma levels of IL-6, IL-8 and IL-10. IL-6 predicted development of MODS with an overall accuracy of 84.7% (specificity: 98.3%, sensitivity: 16.7%). The threshold value for development of MODS was 761.7 pg/ml and 2176.0 pg/ml for mortality during the in patient time.ConclusionWe conclude that plasma IL-6 levels predict mortality and that they are a useful tool to identify patients who are at risk for development of MODS.

Introduction of e-learning in dental radiology reveals significantly improved results in final examination.

PURPOSE Because a traditionally instructed dental radiology lecture course is very time-consuming and labour-intensive, online courseware, including an interactive-learning module, was implemented to support the lectures. The purpose of this study was to evaluate the perceptions of students who have worked with web-based courseware as well as the effect on their results in final examinations. MATERIALS AND METHODS Users (n(3+4)=138) had access to the e-program from any networked computer at any time. Two groups (n(3)=71, n(4)=67) had to pass a final exam after using the e-course. Results were compared with two groups (n(1)=42, n(2)=48) who had studied the same content by attending traditional lectures. In addition a survey of the students was statistically evaluated. RESULTS Most of the respondents reported a positive attitude towards e-learning and would have appreciated more access to computer-assisted instruction. Two years after initiating the e-course the failure rate in the final examination dropped significantly, from 40% to less than 2%. CONCLUSIONS The very positive response to the e-program and improved test scores demonstrated the effectiveness of our e-course as a learning aid. Interactive modules in step with clinical practice provided learning that is not achieved by traditional teaching methods alone. To what extent staff savings are possible is part of a further study.

Biopsy-Controlled Liver Fibrosis Staging Using the Enhanced Liver Fibrosis (ELF) Score Compared to Transient Elastography

Background and Aims Chronic liver diseases are characterized by inflammatory and fibrotic liver injuries that often result in liver cirrhosis with its associated complications such as portal hypertension and hepatocellular carcinoma. Liver biopsy still represents the reference standard for fibrosis staging, although transient elastography is increasingly used for non-invasive monitoring of fibrosis progression. However, this method is not generally available and is associated with technical limitations emphasizing the need for serological biomarkers staging of liver fibrosis. The enhanced liver fibrosis (ELF) score was shown to accurately predict significant liver fibrosis in different liver diseases, although extracellular matrix components detected by this score may not only mirror the extent of liver fibrosis but also inflammatory processes. Methods In this prospective biopsy-controlled study we evaluated the utility of the ELF score in comparison to transient elastography to predict different stages of fibrosis in 102 patients with chronic liver diseases. Results Both techniques revealed similar area under receiver operating characteristic curve values for prediction of advanced fibrosis stages. Compared to transient elastography, the ELF score showed a broader overlap between low and moderate fibrosis stages and a stronger correlation with inflammatory liver injury. Conclusions Both the ELF score as well as transient elastography allowed for high quality fibrosis staging. However, the ELF score was less discriminative in low and moderate fibrosis stages and appeared more strongly influenced by inflammatory liver injury. This should be considered when making clinical interpretations on the basis of ELF score values.

Long-term health-related quality of life after complex and/or complicated esophageal atresia in adults and children registered in a German patient support group

BACKGROUND Health-related quality of life (HRQoL) after esophageal atresia (EA) repair is postulated to be good. However, little is known about the long-term results after repair of complex and/or complicated EA regarding HRQoL. We investigated long-term HRQoL after delayed anastomosis, esophageal replacement, major revisions, or multiple dilatations in patients registered in a support group. METHODS Patients registered in the German patient support group database (KEKS) were enrolled and allocated to subgroups according to surgical treatment and age. HRQoL was evaluated using validated questionnaires (GIQLI, WHO-5, KIDSCREEN27). RESULTS Complete follow-up (mean 14.5 ± 9.8 years) was available for 90/92 patients. Patients were allocated to subgroups delayed anastomosis (n=28), esophageal replacement (n=27), major revisions (n=15), and multiple dilatations (n=20). Adult patients presented with impaired well-being according to WHO-score and gastrointestinal function (GIQLI). In contrast, HRQoL of children was comparable to controls in most KIDSCREEN27-dimensions. Delayed anastomosis was associated with most-favourable HRQoL. Regarding physical well-being, these children scored significantly better than controls [64.01 ± 10.40 vs. 52.36 ± 8.73;p=0.0011], children after replacement [51.40 ± 5.70;p=0.008], revisions [52.04 ± 6.97;p=0.026], and multiple dilatations [50.22 ± 9.67,p=0.04]. CONCLUSIONS HRQoL after complex and/or complicated EA is excellent in children registered in a patient support group. In adults, disease-specific symptoms negatively affect HRQoL. Our data indicate that saving the esophagus may achieve the best HRQoL.

The influence of lunar cycle on frequency of birth, birth complications, neonatal outcome and the gender: A retrospective analysis

Objective. The relation of lunar phases to the birth rate has been the focus of considerable research with still controversial results. The purpose of this study was to examine the influence of the lunar cycle on the frequency of birth, on instrumental delivery, neonatal outcome as well as a possible relationship between lunar position at time of conception and the gender of the child. Material and methods. Six thousand seven hundred and twenty five deliveries were retrospectively analyzed from 01 January 2000 to 31 December 2006 at the Department of Obstetrics and Gynaecology of the University Medical School of Hannover. Inclusion criteria were: spontaneous vaginal births without induction, vacuum and forceps extractions as well as non‐elective cesarean sections, all cases without labor induction. Exact lunar phases were determined by means of birthday and birth time by a specific computer software. Results. The analysis revealed no significant correlation of birth rate to lunar phases. There were also no significant differences regarding birth complications, neonatal outcome and gender to the lunar cycle at the time of conception. Conclusion. There is no predictable influence of the lunar cycle on deliveries or delivery complications nor to the gender of the baby. As expected and in agreement with some recent studies this pervasive myth is not evidence based.

Branched chain amino acids as a parameter for catabolism in treated phenylketonuria

Summary.This study was performed to study an association between nutritional status on one hand and BCAA- and Phe-concentrations on the other hand in PKU patients free of infection. AA profiles from 70 PKU patients were measured. 9 patients (subgroup I) with elevated Phe- and BCAA-concentrations as well as 23 patients (subgroup II) with only elevated Phe-levels were included. Dietary records were obtained from both groups; low caloric intake in subgroup I was increased with Duocal® or p-am ANAMIX® without modifying total protein- and Phe-intake. AA profiles were controlled after 2 weeks.Additionally, we investigated AA profiles from 26 liver transplanted patients with increased carbohydrate and caloric intake as an example for anabolism.In subgroup I Phe- and Isoleu-concentrations decreased sign. after dietary intervention. Leu, Val and Tyr levels decreased not sign. Initial Phe-levels correlated negatively with protein and caloric intake.BCAA concentrations of liver transplanted patients receiving high amounts of carbohydrates were in the lower range of normal.Increased caloric intake lowered most of the elevated Phe- and BCAA- concentrations.

Quality assured ultrasound simulator training for the detection of fetal malformations

Objective. Sonographic training in obstetrics differs broadly in Germany, although there are clearly defined quality‐oriented requirements. In order to improve professional education, a training concept was devised utilizing an ultrasound simulator system. Design. Between October 2004 and May 2006, 100 obstetric ultrasound training courses were held in 12 federal states of Germany. In these daily courses, doctors were trained in the detection of the most common malformations. Sample. One hundred training courses with a total of 1,266 participants. Methods. As a measure of quality assurance, a standardized questionnaire focusing on testing sonographic proficiency before and after the courses was issued in order to analyze the effect of these simulator‐based ultrasound courses. Main outcome measures. Effectiveness of the method with reference to its potential role in structured sonographic training. Results. The concept found prevailing approval (90%) at the level of principle, practical implementation, and clinical usefulness. Of the participants, 91% estimated their subjective training effect as good. The questionnaire analysis showed significant improvement. On average, 75.3% of the questions relating to sonographic proficiency were answered correctly at the end of the course as opposed to 48.6% at the beginning. Conclusion. Structured ultrasound training courses based on an ultrasound simulator system seem to be useful for defining a basic standardized quality of training and significantly improving examiners’ skills. This is a suitable additional instrument to improve the education in obstetric ultrasound.

Partial Rescue of the KIT-Deficient Testicular Phenotype in KitW-v/KitW-v Tg(TSPY) Mice

TSPY encodes the testis-specific protein Y-linked. In man, expression of TSPY is restricted to the testis, where TSPY is expressed in spermatogonia, primary spermatocytes, and round spermatids, and to the prostate gland. There is circumstantial evidence that TSPY is involved in spermatogonial proliferation and gonadal tumorigenesis. Because the laboratory mouse carries the Tspy gene in a naturally silenced state (Tspy-ps), we previously restored TSPY activity in mice and generated a TSPY transgenic mouse line in which the organization and expression of the human TSPY transgene follow the human pattern. In the present study, we generated TSPY transgenic KIT-deficient KitW‑v/KitW‑v mice and analyzed the histology of the testes and epididymides in order to contribute to understanding TSPY function in early germ cell development and spermatogenesis. The KIT receptor and its ligand KITL, previously called stem cell factor, have an indispensable role in hematopoiesis, melanogenesis, and gametogenesis. Homozygous KitW-v mutant male mice on a C57BL/6J background with a mutation in the Kit gene are infertile due to an almost total loss of germ cells in the testes. In this study, histological analyses of testes and epididymides showed an increased number of meiotic and postmeiotic germ cells in KitW‑v/KitW‑v Tg(TSPY) mice compared with age-matched KitW‑v/KitW‑v controls. TSPY was able to restore fertility of some but not all TSPY transgenic KitW‑v/KitW‑v males. Our findings show that TSPY is able to partially rescue spermatogenesis and fertility of KitW‑v/KitW‑v mutants and thereby point to a putative role of TSPY in fetal and adult germ cell proliferation.

TSPY Expression Is Variably Altered in Transgenic Mice with Testicular Feminization

Abstract TSPY (testis-specific protein, Y-encoded) genes are expressed in premeiotic germ cells and round spermatids. The topology and timing of TSPY expression, and also its homology to members of the TTSN-family, suggest that TSPY is a proliferation factor for germ cells. There is also evidence for a role of TSPY in the aetiology of testis cancer. TSPY is a candidate for GBY, the elusive gonadoblastoma locus on the human Y chromosome, which is thought to predispose dysgenetic gonads of 46, XY sex-reversed females to develop gonadoblastoma. We have previously generated a TSPY transgenic mouse line (Tg(TSPY)9Jshm) that carries approximately 50 copies of the human TSPY gene on the mouse Y chromosome. In order to elucidate TSPY expression under complete androgen insensitivity and to investigate a possible role of TSPY in gonadal tumorigenesis, we have now generated sex-reversed TSPY transgenic ArTfm mice hemizygous for the X-linked testicular feminization mutation (ArTfm). We can show that the TSPY transcript is aberrantly spliced in the testes of TSPY-ArTfm mice, and that TSPY expression is upregulated by androgen insensitivity in some but not all animals. TSPY transgenic mice showed significantly increased testes weights. In one TSPY transgenic ArTfm animal, spermatogenesis proceeded beyond meiotic prophase. No tumors of germ cell origin were found in the testes of TSPY-ArTfm mice. Five out of 46 TSPY transgenic ArTfm mice, and 3 out of 31 age-related NMRI-ArTfm controls developed Leydig cell tumors, whereas none of the age-matched ArTfm mice (n = 44) on a wild type background were affected by Leydig cell tumorigenesis.