Brainstem Dysfunction in Healthy Aging.

The brainstem controls sub-cortical and cortical activity and influences the processing of incoming information. The goal of this study was to characterize age related alterations of brainstem-brain interactions during different brain states detected by dynamic analysis of task-free fMRI. 79 young (20-40 years) and 51 older adults (55-80 years) were studied. Internal brainstem structures were segmented using a new multi-contrast segmentation approach. Brain and brainstem gray matter segmentations were warped onto a population template. The ICV-corrected Jacobian determinants were converted into z-score maps and the means from 420 cortical/subcortical/brainstem rois extracted. The fMRI was preprocessed in SPM12/Conn18 and the BOLD signal from 420 cortical/subcortical/brainstem rois extracted. A dynamic task-free analysis approach based on hierarchical cluster analysis was used to identify 15 brain states that were characterized using graph analysis (strength, diversity, modularity). Kruskal-Wallis tests and Spearman correlations were used for statistical analysis. One brain state (cluster 21) occurred more often in older adults (p=0.008). It was characterized by a lower mean modular strength and brainstem-cortical strength in older adults compared to younger adults. Global age related gray matter differences were positively correlated with brain state 21's modular strength. Furthermore, brain state 21 duration was negatively correlated with working memory (r = -0.28, p=0.002). The findings suggest an age related weakening of the within and between network synchronization at the brainstem level during brain state 21 in older adults that negatively affects cortical and subcortical synchronization and working memory performance.

Interferon alfa-2a maintenance after salvage autologous stem cell transplantation in atypical mycosis fungoides with central nervous system involvement.

Mycosis fungoides (MF) is a primary cutaneous T-cell lymphoma with unfavourable prognosis for patients with advanced stages of the disease. Refractory disease and advanced-stage disease require systemic therapy. We report on a rare case of an atypical predominantly CD8+ folliculotropic MF, a subtype of MF with poorer prognosis, in a 59-year-old woman. She was initially diagnosed with MF restricted to the skin, of T3N0M0B0/stage IIB according to the current World Health Organization-European Organisation for Research and Treatment of Cancer classification. First-line treatment with local percutaneous radiotherapy in combination with systemic interferon alfa-2a resulted in complete remission. However, 21 months later the disease progressed to T3N0M1B0/stage IVB with development of cerebral manifestation and thus very poor prognosis. Allogeneic stem cell transplantation (SCT) was not a therapeutic option due to the lack of a suitable donor. We initiated methotrexate and cytarabine chemotherapy, followed by high-dose chemotherapy with thiotepa and carmustine with autologous SCT. Despite rapid response and complete remission of the cerebral lesions, disease recurrence of the skin occurred soon after. Interestingly, readministration of interferon alfa-2a as a maintenance treatment after the salvage autologous SCT resulted in a durable complete remission during the follow-up period of currently 17 months after autologous SCT. What's already known about this topic? Mycosis fungoides is a primary cutaneous T-cell lymphoma with unfavourable prognosis for the advanced stages of the disease. A refractory course of disease requires systemic therapy. What does this study add? We report on an unusual case of a patient with mycosis fungoides with cerebral involvement, in which a durable complete remission was achieved upon autologous stem cell therapy and interferon alfa-2a maintenance therapy.

[Importance of investigation of fetal eyes : Supplement to fetal autopsy]. / Bedeutung der Untersuchung fetaler Augen : Ergänzung der Fetobduktion.

The investigation of fetal eyes is a subspecialty, which is performed by only a few ophthalmic pathologists and pathologists in specialized centers. If a certain fetal syndrome is suspected, the fetal eyes should be removed and submitted for ophthalmic pathological investigation in a specialized center. This can provide additional diagnostic information allowing a final classification of a syndrome and the comprehensive genetic counselling of parents with respect to future pregnancies. This article provides an overview of the histopathological findings in fetal eyes, which are of particular relevance for the pediatric pathological autopsy. In addition, the basic points of ocular development, the preparation of fetal eyes and frequent artifacts are presented.

Pcgf6, a polycomb group protein, regulates mesodermal lineage differentiation in murine ESCs and functions in iPS reprogramming.

Polycomb group (PcG) proteins comprise evolutionary conserved factors with essential functions for embryonic development and adult stem cells. PcG proteins constitute two main multiprotein polycomb repressive complexes (PRC1 and PRC2) that operate in a hierarchical manner to silence gene transcription. Functionally distinct PRC1 complexes are defined by Polycomb group RING finger protein (Pcgf) paralogs. So far, six Pcgf paralogs (Pcgf1-6) have been identified as defining components of different PCR1-type complexes. Paralog-specific functions are not well understood. Here, we show that Pcgf6 is the only Pcgf paralog with high expression in undifferentiated embryonic stem cells (ESCs). Upon differentiation Pcgf6 expression declines. Following Pcgf6 kockdown (KD) in ESCs, the expression of pluripotency genes decreased, while mesodermal- and spermatogenesis-specific genes were derepressed. Concomitantly with the elevated expression of mesodermal lineage markers, Pcgf6 KD ESCs showed increased hemangioblastic and hematopoietic activities upon differentiation suggesting a function of Pcgf6 in repressing mesodermal-specific lineage genes. Consistant with a role in pluripotency, Pcgf6 replaced Sox2 in the generation of germline-competent induced pluripotent stem (iPS) cells. Furthermore, Pcgf6 KD in mouse embryonic fibroblasts reduced the formation of ESC-like colonies in OSKM-driven reprogramming. Together, these analyses indicate that Pcgf6 is nonredundantly involved in maintaining the pluripotent nature of ESCs and it functions in iPS reprogramming.

[Stillbirths in Germany: Retrospective Analysis of 168 Cases between 2003 and 2011]. / Totgeburten in Deutschland: Retrospektive Datenanalyse von 168 Fällen zwischen 2003 und 2011.

BACKGROUND: The decline in the incidence of stillbirths in Germany has remained static in recent years. This study aims to analyse the current situation of data documentation and examination of stillbirths. Furthermore, possible stillbirth prevention strategies should be developed. METHODS: Searches in the international peer-reviewed literature, retrospective data collection of 168 stillbirths in 8 hospitals, (in the area of Bonn) with subsequent statistical evaluation (descriptive statistics, t-test and binominal test) were undertaken. RESULTS: This study shows considerable deficits in data documentation, interdisciplinary communication and postmortal examination. Only in 51.8% (87/168) of the cases was a certain or uncertain cause of death found (42.3% placental, 1.2% foetal, 3.6% chromosomal, 4.8% umbilical cord abnormalities). Severe foetal growth restriction (<5(th) percentile) was observed in 29.2%; 44.9% (22/49) of them died at the age of ≥36+0 weeks of gestation. CONCLUSION: The first step to reduce the rate of stillbirths in Germany is to increase the identified causes of foetal death: Therefore, an interdisciplinary case report form was compiled to improve data collection and interdisciplinary collaboration. To standardise and complete postmortal management, an algorithm was created. The long-term aim is the development of a central data register for statistical analysis, to identify goals of research and to organise conferences with interdisciplinary reports of diagnostic findings.

[Massive ovarian edema in a 13-year-old girl]. / Massives Ovarialödem bei einem 13-jährigen Mädchen.

Massive ovarian edema is a rare tumor-like condition found in young women resulting from accumulation of fluid mostly due to partial or intermittent torsion of the ovary or secondary, to a preexisting ovarian lesion. We report a case of a 13-year-old girl presenting with an ovarian mass measuring 16 cm. CA-12-5 levels were slightly elevated. Concerns regarding underlying malignancy led to salpingo-oophorectomy. Pathological evaluation revealed a massive ovarian edema and multiple thromboses of ovarian veins. Differentiating massive ovarian edema from malignant ovarian tumor is crucial to prevent patients from undergoing unnecessary surgery.

Post-discharge mortality in adult patients hospitalized for tuberculosis: a prospective cohort study.

Determining outcomes and predictors of mortality following discharge from tuberculosis (TB) hospitalization is crucial to establish health policies. The objective of this study was to analyze outcomes and, secondarily, predictors of mortality following discharge from TB hospitalization. This was a prospective cohort study of patients diagnosed with TB (all forms) discharged from the hospital who began treatment during hospitalization. Out of 169 subjects included, 38 died during the 13-months of follow-up, within a median of 3 months (95%CI: 1.49-4.51). In the multivariate analysis, the variables independently associated with death were age (HR=1.04, 95%CI: 1.01-1.06, P=0.001), presence of sputum production (HR=2.18, 95%CI: 1.09-4.34, P=0.027), and Charlson Comorbidity Index (HR=1.19, 95%CI: 1.04-1.36, P=0.015). In conclusion, post-discharge mortality in subjects hospitalized for TB was 22.5%, with mean survival of 4.6 months. The mortality was higher in older subjects, in those who reported sputum production, and in those with a high comorbidity index.

Tissue engineering approaches to degenerative disc disease--a meta-analysis of controlled animal trials.

OBJECTIVE: The objective of this systematic review was to assess cell/biomaterial treatments of degenerative disc disease in controlled animal trails. The primary endpoints were restoration of disc height and T2 signal intensity. METHOD: PubMed, CINAHL, EMBASE, Cochrane Central Register of Controlled Trials (CENTRAL), and Cochrane Database of Systematic Reviews (CDSR) were searched for studies reporting on the use of tissue engineering treatments (cells/biomaterials/cells and biomaterials) for degenerative disc disease treatments in a controlled trial. Publication bias was assessed graphically using funnel plots and Egger's regression. Data were grouped by follow-up duration - early (<4 weeks), intermediate (4-12 weeks) and late (>12 weeks), and weighted mean differences (WMD) were calculated using DerSimonian-Laird Random Effect models. RESULTS: Thirteen papers, published between 2004 and 2011, were included in this study. In comparison with the injured disc, all three treatments showed a positive effect in disc height, but none of the treatments restored disc height compared to the healthy disc. Overall, there seemed to be a better effect on disc height restoration for the treatment with cells and biomaterials. None of the treatments could achieve the same T2 signal intensity as the healthy disc, and compared to the injured disc, only the treatment with cells and biomaterials showed consistently better results. CONCLUSION: Treatment of an injured/degenerating disc with cells, cells plus biomaterial or biomaterial alone has a potential for at least a partial regeneration of the disc. However, so far, none of the treatments is able to effectively restore the properties of a healthy disc.

Upper abdominal cysts in first trimester fetuses.

Cystic abdominal structures in first trimester fetuses are rare. In order to elucidate the clinical significance of first trimester abdominal cysts, we present three cases of the ultrasonographic detection of upper abdominal cysts in first trimester fetuses with spontaneous regression until birth. Cysts with maximum diameters of 20, 15 and 21 mm were diagnosed in the first scan at 12 + 3 weeks of gestation and two times at 13 + 0 weeks of gestation, respectively. They had echogenic walls, a longish shape and were all located in the upper part of the abdomen. Further anomalies, relevant maternal infections, the most frequent cystic fibrosis mutations and fetal chromosomal abnormalities were excluded. In all three cases follow-up scans showed similar characteristics: The relatively large intraabdominal cyst resolved during the early second trimester. A hyperdense structure of up to 24 mm close to the liver capsule was the sole prenatally detectable remnant. During the first months after birth, development of all three children was normal. Pediatric ultrasound examination reported subcapsular liver calcifications. In conclusion, these cases demonstrate that first trimester upper abdominal cysts with spontaneous resolution until birth are associated with a favorable outcome, if infections and additional anatomical or chromosomal anomalies are ruled out. Nevertheless, one should be aware that postnatal gastrointestinal complications have been described in a few cases, even if the cyst had resolved spontaneously in utero.

[Congenital heart defects of the septa, endocardial cushions and the conotruncus]. / Angeborene Herzfehlbildungen von Septen, Endokardkissen und Konotrunkus.

During embryological development the heart develops from a simple tube into a complex fully developed heart with four chambers. Hence all congenital heart defects develop before the ninth week of gestation. Currently a steadily increasing number of genetic mutations have been found to be responsible for congenital heart defects. Nevertheless, up to now it has been impossible to diagnose a heart defect just on the basis of molecular pathology. Despite the current excellent prenatal and postnatal ultrasound diagnostics, the post-mortem examination is still the gold standard for the diagnosis of complex heart malformations. However, this requires knowledge of the pathomorphology of the heart malformation in question. Therefore, characteristic and distinguishing features of septal defects including atrioventricular septal defects are presented, especially as the latter are part of complex heart defects, such as conotruncal heart malformations.

Analysis of copy number variation in men with non-obstructive azoospermia.

BACKGROUND: Recent findings demonstrate that single nucleotide variants can cause non-obstructive azoospermia (NOA). In contrast, copy number variants (CNVs) were only analysed in few studies in infertile men. Some have reported a higher prevalence of CNVs in infertile versus fertile men. OBJECTIVES: This study aimed to elucidate if CNVs are associated with NOA. MATERIALS AND METHODS: We performed array-based comparative genomic hybridisation (aCGH) in 37 men with meiotic arrest, 194 men with Sertoli cell-only phenotype, and 21 control men. We filtered our data for deletions affecting genes and prioritised the affected genes according to the literature search. Prevalence of CNVs was compared between all groups. Exome data of 2,030 men were screened to detect further genetic variants in prioritised genes. Modelling was performed for the protein encoded by the novel candidate gene TEKT5 and we stained for TEKT5 in human testicular tissue. RESULTS: We determined the cause of infertility in two individuals with homozygous deletions of SYCE1 and in one individual with a heterozygous deletion of SYCE1 combined with a likely pathogenic missense variant on the second allele. We detected heterozygous deletions affecting MLH3, EIF2B2, SLX4, CLPP and TEKT5, in one subject each. CNVs were not detected more frequently in infertile men compared with controls. DISCUSSION: While SYCE1 and MLH3 encode known meiosis-specific proteins, much less is known about the proteins encoded by the other identified candidate genes, warranting further analyses. We were able to identify the cause of infertility in one out of the 231 infertile men by aCGH and in two men by using exome sequencing data. CONCLUSION: As aCGH and exome sequencing are both expensive methods, combining both in a clinical routine is not an effective strategy. Instead, using CNV calling from exome data has recently become more precise, potentially making aCGH dispensable.

Ebstein's anomaly of the tricuspid valve in association with tetralogy of fallot and absent pulmonary valve syndrome.

Ebstein's anomaly and absent pulmonary valve syndrome belong to the rarest congenital heart defects. Their association has never been reported so far. We present the unusual case of a fetus at 23 weeks of gestation with Ebstein's anomaly, tetralogy of Fallot, absent pulmonary valve and agenesis of the arterial duct. The main diagnostic features were apical displacement of the septal leaflet of the tricuspid valve with an offset from the mitral valve of 8 mm, a pronounced atrialization of the right ventricle, a large malalignment ventricular septal defect with overriding aorta in combination with absence of the pulmonary valve leaflets, to and fro flow pattern over the stenotic pulmonary valve annulus, turbulent flow in the pulmonary trunk, massive dilatation of the pulmonary trunk plus the pulmonary arteries and a right aortic arch with retroesophageal course of an aberrant left subclavian artery. The arterial duct and the thymus were absent. The remaining fetal anatomy was unremarkable. Amniocentesis revealed a normal male karyotype; 22q11 microdeletion was ruled out. After being counseled on the unfavorable prognosis, the parents opted for termination of pregnancy. The prenatal cardiac findings were confirmed at autopsy; however, a severely hypoplastic thymus was found instead of the suspected aplasia.

Towards an intraoperative engineering of osteogenic and vasculogenic grafts from the stromal vascular fraction of human adipose tissue.

Grafts generated by cultivation of progenitor cells from the stromal vascular fraction of human adipose tissue have been proven to have osteogenic and vasculogenic properties in vivo. However, in vitro manufacture of such implants is challenged by complex, impractical and expensive processes, and requires implantation in a separate surgery. This study investigates the feasibility of an intraoperative approach to engineer cell-based bone grafts with tissue harvest, cell isolation, cell seeding onto a scaffold and subsequent implantation within a few hours. Freshly isolated adipose tissue cells from a total of 11 donors, containing variable fractions of mesenchymal and endothelial progenitors, were embedded at different densities in a fibrin hydrogel, which was wrapped around bone substitute materials based on beta-tricalcium phosphate (ChronOS), hydroxyapatite (Engipore), or acellular xenograft (Bio-Oss). The resulting constructs, generated within 3 hours from biopsy harvest, were immediately implanted ectopically in nude mice and analysed after eight weeks. All explants contained blood vessels formed by human endothelial cells, functionally connected to the recipient's vasculature. Human origin cells were also found within osteoid structures, positively immunostained for bone sialoprotein and osteocalcin. However, even with the highest loaded cell densities, no frank bone tissue was detected, independently of the material used. These results provide a proof-of-principle that an intraoperative engineering of autologous cell-based vasculogenic bone substitutes is feasible, but highlight that - in the absence of in vitro commitment--additional cues (e.g., low dose of osteogenic factors or orthotopic environmental conditions) are likely needed to support complete osteoblastic cell differentiation and bone tissue generation.

[Appendicitis in childhood: correlation of clinical data with histopathological findings]. / Akute Appendizitis im Kindesalter: Korrelation klinischer und histopathologischer Befunde.

BACKGROUND: Quantifying the histopathological diagnoses of appendectomies in daily routine paidopathology results in a high percentage of appendices without histomorphological sign of acute inflammation. To identify clinical factors significantly associated with the morphological diagnosis, histopathological findings and clinical data--documented in patients' files--were examined. PATIENTS: All 856 children (age: 5 m-15 yrs) whose appendix had been resected within a 7-year-period were--depending on the histopathological diagnoses--allocated to the group "appendix without" resp. "appendix with signs of acute inflammation". METHOD: All files were examined concerning anamnestic data, clinical signs of acute appendicitis and laboratory parameters. The data were analysed by χ(2)-test and Wilcoxon-test concerning differences between the 2 groups with regard to the anamnestic and clinical facts and parameters. Using binary logistic regression, these clinical parameters were analyzed in correlation with the histopathological diagnoses. RESULTS: By consideration of the factors "leucocyte count", "vomiting" and "percussion tenderness" 75% of the children would have been allocated to the accurate postoperative pathomorphological diagnosis. CONCLUSIONS: There was a significant correlation of "leucocyte count", "vomiting" and "percussion tenderness" with histopathology in 75% of the children. As hence 25% were not allocated correctly this combination is no unequivocal combination for prediction or exclusion of an acute appendicitis. Hence, even with these statistically significant parameters a comparably high percentage of falsely-positive appendectomies has to be anticipated.

Renal cell carcinoma with Xp11.2 translocation in a 7-year-old boy.

BACKGROUND: More than 90% of pediatric renal tumors are nephroblastomas while renal cell carcinomas (RCC) are rare in children (< 5%). PATIENT: According to the clinical diagnoses of a nephroblastoma stage IV a 7-year-old boy with a kidney tumor and peripheral pulmonary lesion was preoperatively treated for 8 weeks with Vincristine, Actinomycin D and Adriamycin. The resected kidney displayed a RCC with Xp11.2 translocation. There was no tumor regression and the pulmonary lesion was no longer detectable. Hence chemotherapy was put to a halt. CONCLUSION: Fine needle aspiration biopsy (FNA) would have allowed to adjust the tumor subtype. Prognosis of pediatric RCC with translocation seems more favourable than without translocation though definitive evidence will only be possible by documentation in a clinical diagnose-related register.

Preparing healthcare facilities to operate safely and effectively during the COVID-19 pandemic: The missing piece in the puzzle.

The stated objective of the COVID-19 lockdown was to allow time to prepare healthcare facilities. Preparation must include administrative and environmental measures, which when combined with personal protective equipment, minimise the risk of the spread of infection to patients and healthcare workers (HCWs) in facilities, allowing HCWs to safely provide essential services during the pandemic and limit the indirect effects of COVID-19 caused by healthcare disruption. We present our model for facility preparation based on colour-coded zones, social distancing, hand hygiene, rapid triage and separate management of symptomatic patients, and attention to infection transmission prevention between HCWs in communal staff areas. This model specifically addresses the challenges in preparing a facility for COVID-19 in a low-resource setting and in rural areas. In addition, we include links to resources to allow workers in low-resource settings to prepare their facilities adequately.