Aberrant Promoter CpG Methylation Is a Mechanism for Impaired PHD3 Expression in a Diverse Set of Malignant Cells

The prolyl-hydroxylase domain family of enzymes (PHD1-3) plays an important role in the cellular response to hypoxia by negatively regulating HIF-α proteins. Disruption of this process can lead to up-regulation of factors that promote tumorigenesis. We observed decreased basal expression of PHD3 in prostate cancer tissue and tumor cell lines representing diverse tissues of origin. Furthermore, some cancer lines displayed a failure of PHD3 mRNA induction when introduced to a hypoxic environment. This study explores the mechanism by which malignancies neither basally express PHD3 nor induce PHD3 under hypoxic conditions.Using bisulfite sequencing and methylated DNA enrichment procedures, we identified human PHD3 promoter hypermethylation in prostate, breast, melanoma and renal carcinoma cell lines. In contrast, non-transformed human prostate and breast epithelial cell lines contained PHD3 CpG islands that were unmethylated and responded normally to hypoxia by upregulating PHD3 mRNA. Only treatment of cells lines containing PHD3 promoter hypermethylation with the demethylating drug 5-aza-2'-deoxycytidine significantly increased the expression of PHD3.We conclude that expression of PHD3 is silenced by aberrant CpG methylation of the PHD3 promoter in a subset of human carcinoma cell lines of diverse origin and that this aberrant cytosine methylation status is the mechanism by which these cancer cell lines fail to upregulate PHD3 mRNA. We further show that a loss of PHD3 expression does not correlate with an increase in HIF-1α protein levels or an increase in the transcriptional activity of HIF, suggesting that loss of PHD3 may convey a selective advantage in some cancers by affecting pathway(s) other than HIF

Ethical issues in autologous stem cell transplantation (ASCT) in advanced breast cancer: A systematic literature review

BACKGROUND: An effectiveness assessment on ASCT in locally advanced and metastatic breast cancer identified serious ethical issues associated with this intervention. Our objective was to systematically review these aspects by means of a literature analysis. METHODS: We chose the reflexive Socratic approach as the review method using Hofmann's question list, conducted a comprehensive literature search in biomedical, psychological and ethics bibliographic databases and screened the resulting hits in a 2-step selection process. Relevant arguments were assembled from the included articles, and were assessed and assigned to the question list. Hofmann's questions were addressed by synthesizing these arguments. RESULTS: Of the identified 879 documents 102 included arguments related to one or more questions from Hofmann's question list. The most important ethical issues were the implementation of ASCT in clinical practice on the basis of phase-II trials in the 1990s and the publication of falsified data in the first randomized controlled trials (Bezwoda fraud), which caused significant negative effects on recruiting patients for further clinical trials and the doctor-patient relationship. Recent meta-analyses report a marginal effect in prolonging disease-free survival, accompanied by severe harms, including death. ASCT in breast cancer remains a stigmatized technology. Reported health-related-quality-of-life data are often at high risk of bias in favor of the survivors. Furthermore little attention has been paid to those patients who were dying. CONCLUSIONS: The questions were addressed in different degrees of completeness. All arguments were assignable to the questions. The central ethical dimensions of ASCT could be discussed by reviewing the published literature

Bioinformatics and molecular modeling in glycobiology

The field of glycobiology is concerned with the study of the structure, properties, and biological functions of the family of biomolecules called carbohydrates. Bioinformatics for glycobiology is a particularly challenging field, because carbohydrates exhibit a high structural diversity and their chains are often branched. Significant improvements in experimental analytical methods over recent years have led to a tremendous increase in the amount of carbohydrate structure data generated. Consequently, the availability of databases and tools to store, retrieve and analyze these data in an efficient way is of fundamental importance to progress in glycobiology. In this review, the various graphical representations and sequence formats of carbohydrates are introduced, and an overview of newly developed databases, the latest developments in sequence alignment and data mining, and tools to support experimental glycan analysis are presented. Finally, the field of structural glycoinformatics and molecular modeling of carbohydrates, glycoproteins, and protein–carbohydrate interaction are reviewed

Listeria pathogenesis and molecular virulence determinants

The gram-positive bacterium Listeria monocytogenes is the causative agent of listeriosis, a highly fatal opportunistic foodborne infection. Pregnant women, neonates, the elderly, and debilitated or immunocompromised patients in general are predominantly affected, although the disease can also develop in normal individuals. Clinical manifestations of invasive listeriosis are usually severe and include abortion, sepsis, and meningoencephalitis. Listeriosis can also manifest as a febrile gastroenteritis syndrome. In addition to humans, L. monocytogenes affects many vertebrate species, including birds. Listeria ivanovii, a second pathogenic species of the genus, is specific for ruminants. Our current view of the pathophysiology of listeriosis derives largely from studies with the mouse infection model. Pathogenic listeriae enter the host primarily through the intestine. The liver is thought to be their first target organ after intestinal translocation. In the liver, listeriae actively multiply until the infection is controlled by a cell-mediated immune response. This initial, subclinical step of listeriosis is thought to be common due to the frequent presence of pathogenic L. monocytogenes in food. In normal indivuals, the continual exposure to listerial antigens probably contributes to the maintenance of anti-Listeria memory T cells. However, in debilitated and immunocompromised patients, the unrestricted proliferation of listeriae in the liver may result in prolonged low-level bacteremia, leading to invasion of the preferred secondary target organs (the brain and the gravid uterus) and to overt clinical disease. L. monocytogenes and L. ivanovii are facultative intracellular parasites able to survive in macrophages and to invade a variety of normally nonphagocytic cells, such as epithelial cells, hepatocytes, and endothelial cells. In all these cell types, pathogenic listeriae go through an intracellular life cycle involving early escape from the phagocytic vacuole, rapid intracytoplasmic multiplication, bacterially induced actin-based motility, and direct spread to neighboring cells, in which they reinitiate the cycle. In this way, listeriae disseminate in host tissues sheltered from the humoral arm of the immune system. Over the last 15 years, a number of virulence factors involved in key steps of this intracellular life cycle have been identified. This review describes in detail the molecular determinants of Listeria virulence and their mechanism of action and summarizes the current knowledge on the pathophysiology of listeriosis and the cell biology and host cell responses to Listeria infection. This article provides an updated perspective of the development of our understanding of Listeria pathogenesis from the first molecular genetic analyses of virulence mechanisms reported in 1985 until the start of the genomic era of Listeria research

Entwicklung der Morbidität und Mortalität von SARS-CoV-2 in Altenpflegeheimen in Frankfurt am Main, Deutschland, von 2020 bis 2022: Welche Schutzmaßnahmen sind noch erforderlich?

Introduction: Nursing-home residents are among the highest risk group in the SARS-CoV-2 pandemic. At the onset of the SARS-CoV-2 pandemic, the majority of all deaths from or with SARS-CoV-2 occurred in long-term care facilities (LTCFs), so that maximum protective measures were mandated for these facilities. This study analyzed the impact of the new virus variants and the vaccination campaign on disease severity and mortality among nursing home residents and staff through 2022 as a basis for determining which protective measures remain necessary and appropriate. Methods: In five homes in Frankfurt am Main, Germany, with a total capacity for 705 residents, all cases occurring in the facility among residents and staff were recorded and documented (date of birth and diagnosis, hospitalization and death, vaccination status) and were descriptively analyzed with SPSS.Results: By 31st August 2022, 496 residents tested positive for SARS-CoV-2, 93 in 2020, 136 in 2021, and 267 in 2022; 14 residents presented with a second SARS-CoV-2 infection in 2022, having previously experienced an infection in 2020 or 2021. The percentage of hospitalizations decreased from 24.7% (2020) and 17.6% (2021) to 7.5% (2022), and the percentage of deaths decreased from 20.4% and 19.1% to 1.5%. In 2021, 61.8% of those infected were vaccinated (at least 2x); in 2022, 86.2% of residents had been vaccinated twice, 84% of whom had already had a booster vaccination. Hospitalization and death rates were significantly higher among the unvaccinated than the vaccinated throughout all years (unvaccinated 21.5% and 18.0%; vaccinated 9.8% and 5.5%; KW test p=0.000). However, this difference was no longer significant under the prevalence of the Omicron variant in 2022 (unvaccinated 8.3% and 0%; p=0.561; vaccinated 7.4% and 1.7%; p=0.604). From 2020 to 2022, 400 employees were documented as infected, with 25 having second infections in 2022. Only one employee showed a second infection in 2021 following the first in 2020. Three employees were hospitalized; no deaths occurred. Discussion and conclusion: Severe COVID-19 courses occurred with the Wuhan Wild type in 2020, with a high death rate among nursing-home residents. In contrast, during the waves in 2022 with the relatively mildly pathogenic Omicron variant, many infections but few severe courses and deaths were observed among the now mostly vaccinated and boostered nursing-home residents. Given the high immunity of the population and the low pathogenicity of the circulating virus - even in nursing-home residents - protective measures in nursing homes that restrict people's right to self-determination and quality of life no longer seem justified. Instead, the general hygiene rules and the recommendations of the KRINKO (German Commission for Hospital Hygiene and Infection Prevention) on infection prevention should be followed, and the recommendations of the STIKO (German Standing Commission on Vaccination) on vaccination not only against SARS-CoV-2 but also against influenza and pneumococci should be observed.Einleitung: Altenpflegeheimbewohner zählen zu der höchsten Risikogruppe in der SARS-CoV-2-Pandemie. Zu Beginn der SARS-CoV-2-Pandemie traten die Mehrzahl aller Todesfälle an oder mit SARS-CoV-2 in Altenpflegeheimen (APHs) auf, weshalb für die Einrichtungen maximale Schutzmaßnahmen verfügt wurden. Die Auswirkungen der neuen Virus-Varianten und der Impfkampagne auf die Krankheitsschwere und Sterberate bei Altenpflegeheimbewohnern bis 2022 wird untersucht - als Grundlage für die Frage, welche Schutzmaßnahmen weiterhin erforderlich und angemessen sind. Methode: In fünf Heimen in Frankfurt am Main mit insgesamt 705 Bewohnerplätzen wurden alle in der Einrichtung auftretenden Fälle sowohl bei Bewohnern als auch bei Mitarbeitern erfasst und dokumentiert (Geburts- und Diagnosedatum, Hospitalisierung und Tod, Impfstatus) und mit SPSS deskriptiv ausgewertet.Ergebnisse: Bis August 2022 wurden 496 Bewohner positiv auf SARS-CoV-2 getestet, 93 im Jahr 2020, 136 im Jahr 2021 und 267 im Jahr 2022. 14 Bewohner wiesen im Jahr 2022 eine zweite SARS-CoV-2-Infektion auf, nachdem sie bereits 2020 oder 2021 eine Infektion durchgemacht hatten. Der Anteil der Hospitalisierungen nahm von 24,7% (2020) über 17,6% (2021) auf 7,5% (2022) ab, der Anteil der Todesfälle von 20,4% über 19,1% auf 1,5%. Im Jahr 2021 waren 61,8% der Infizierten geimpft (mindestens 2x), im Jahr 2022 waren 86,2% der Bewohner geimpft, 84% davon bereits "geboostert". Die Hospitalisierungs- und Todesrate war bei den Ungeimpften in der Gesamtbetrachtung signifikant höher als bei den Geimpften (Ungeimpfte 21,5% und 18,0%; Geimpfte 9,8% und 5,5%; KW Test p=0,000), allerdings war dieser Unterschied unter dem Vorherrschen der Omikron-Variante im Jahr 2022 nicht mehr signifikant (Ungeimpfte 8,3% und 0%; p=0,561; Geimpfte 7,4% und 1,7%; p=0,604). Von 2020 bis 2022 wurden 400 Mitarbeiter als infiziert dokumentiert, 25 Mitarbeiter wiesen eine Zweitinfektion im Jahr 2022 auf. Nur bei einem Mitarbeiter war die zweite Infektion bereits 2021 aufgetreten, nach der ersten im Jahr 2020. Drei Mitarbeiter wurden hospitalisiert, Todesfälle traten nicht auf. Diskussion und Schlussfolgerung: 2020 kam es unter dem Wuhan-Wild-Typ zu schweren COVID-19-Verläufen mit einer hohen Todesrate unter den Altenpflegeheimbewohnern. Demgegenüber waren während der Wellen in 2022 mit der wenig pathogenen Omicron-Variante bei den inzwischen zumeist geimpften und geboosterten Altenpflegeheimbewohnern zwar viele Infektionen aber nur wenige schwere Verläufe und Todesfälle zu beobachten. Angesichts der hohen Immunität der Bevölkerung und der geringen Pathogenität des zirkulierenden Virus - auch bei Altenpflegeheimbewohnern - erscheinen. Schutzmaßnahmen in Altenpflegeheimen, die das Selbstbestimmungsrecht und die Lebensqualität der Menschen einschränken, nicht mehr gerechtfertigt. Stattdessen sollten die allgemeinen Hygieneregeln und die Empfehlungen der KRINKO (Kommission für Krankenhaus-hygiene und Infektionsprävention) zur Infektionsprävention umgesetzt sowie die Empfehlungen der STIKO (Ständige Impfkommission) zur Impfung nicht nur gegen SARS-CoV-2 sondern auch gegen Influenza und Pneumokokken beachtet werden