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For the first time in medical history, the result of a laboratory assay (RT-qPCR) was used as the sole criterion to diagnose a disease (Covid-19) without any rating of clinical symptoms and to justify the implementation of population-wide non-pharmaceutical interventions during virus season. The aim of this study is (1) to evaluate a robust RT-qPCR test that overcomes major concerns raised within the scientific community on the first WHO-recommended RT-qPCR protocol to detect SARS-CoV-2 sequences, (2) to genomically characterize individual SARS-CoV-2 strains circulating in the Czech Republic from autumn 2020 to spring 2021 applying NGS and (3) to re-initiate the currently missing scientific dialogue and return to reason and evidence-based medicine. We present a RT-qPCR test designed for the detection of all SARS-CoV-2 variants known so far. Based on the genomic mutation profile, we demonstrate that the three individual waves culminating from autumn 2020 to spring 2021 in the Czech Republic were successive, but lacked direct genomic relationship between each other. This became obvious with the omicron variant, which did not reveal direct evolutionary connection to any of the previous SARS-CoV-2 variants. In addition, we provide evidence that neglected principles of good scientific practice resulted not only in the publication of the WHO-recommended Charité RT-qPCR protocol, but also in health-related problems. Unnecessary quarantine of false positive-tested, healthy individuals, as well as lockdowns and atrocious collateral damage on societies and economies worldwide due to a high number of false-positive “PCR-cases.” Further, some chain of infection caused by false negative-tested, symptomatic individuals lead to real Covid-19 clusters. Both our results and literature data confirm that validation of any PCR-based diagnostic test by sequencing is mandatory on a regular basis. To prevent future misconduct, science needs a reality check and must re-initiate the scientific dialogue and liberate itself from political influence and dogma.

ZusammenfassungEs gibt bislang nur wenige Studien, die den Einfluss einer streng kohlenhydratreduzierten, sehr fettreichen, sog. „ketogenen“ Kost auf den Verlauf einer Krebserkrankung untersuchten. Da eine solche Kostform längerfristig Schwierigkeiten bei der Compliance zu Hause bereitet, ist ein supportiver Therapieansatz evtl. sinnvoller, weil er in Anlehnung an Fastenkuren mit den oxidativen Krebstherapien kombiniert werden kann.

Increased intestinal permeability and inflammation, both fueled by dysbiosis, appear to contribute to rheumatoid arthritis (RA) pathogenesis. This single-center pilot study aimed to investigate zonulin, a marker of intestinal permeability, and calprotectin, a marker of intestinal inflammation, measured in serum and fecal samples of RA patients using commercially available kits. We also analyzed plasma lipopolysaccharide (LPS) levels, a marker of intestinal permeability and inflammation. Furthermore, univariate, and multivariate regression analyses were carried out to determine whether or not there were associations of zonulin and calprotectin with LPS, BMI, gender, age, RA-specific parameters, fiber intake, and short-chain fatty acids in the gut. Serum zonulin levels were more likely to be abnormal with a longer disease duration and fecal zonulin levels were inversely associated with age. A strong association between fecal and serum calprotectin and between fecal calprotectin and LPS were found in males, but not in females, independent of other biomarkers, suggesting that fecal calprotectin may be a more specific biomarker than serum calprotectin is of intestinal inflammation in RA. Since this was a proof-of-principle study without a healthy control group, further research is needed to validate fecal and serum zonulin as valid biomarkers of RA in comparison with other promising biomarkers.

Dietary factors probably play a role in the pathogenesis and clinical course of rheumatoid arthritis (RA). There is a paucity of specific dietary guidelines for RA patients and little information on their implementation in daily life. Therefore, this study aimed to determine the nutritional status and provision of nutritional education among outpatients with RA. Here, 61 patients were included with a sex ratio of 2.03 (f/m). Based on BMI, 22% of women were overweight and 32% obese, whereas 50% of men were overweight and 30% obese. Fasting blood and a 3-day estimated dietary record were collected. Additionally, patients were asked whether they had already received information about a specific diet as part of their disease treatment plan. Elevated total cholesterol levels were found in 76% of women and in 60% of men caused by increased non-HDL-C levels. The dietary intake assessment showed a lower self-reported intake of energy, polyunsaturated fat, carbohydrates, fiber, and several micronutrients than recommended. Regarding healthy eating, all patients reported familiarity with dietary recommendations, but found it difficult to implement the recommendations into their diets. These findings suggested that RA patients need more specific recommendations and education in clinical practice to improve the quality of their diet.

Background/Objectives: BRCA mutation carriers and women at high risk of breast/ovarian cancer are faced with the intricate question to opt for prophylactic surgeries and/or a periodic screening. The aim of this study was therefore to identify objective and emotional factors that have an impact on the decision-making process. Methods: Ninety-five women with BRCA mutations or women at increased breast/ovarian cancer lifetime risk were counseled at our outpatient department and either opted for prophylactic surgery or periodic screening. To identify the psychological factors that could have influenced the decision-making, a standardized questionnaire was applied. Additionally, clinical data were collected and were reviewed by a personal talk. Results: Seventy-one of the patients opted for an increased surveillance only, 21 for prophylactic surgeries. Positive predictors for prophylactic surgeries were sociodemographic characteristics such as parity and objective variables such as verified mutation status. Hierarchical regression analysis revealed that the need for safety in health issues has been the only significant psychological predictor of surgery beyond the objective factors. Fear of surgical procedures, menopausal symptoms after surgery, loss of attractiveness, or fear of interferences with sexual life did not significantly affect decision-making. Conclusion: Decision-making towards prophylactic surgeries is influenced by objective but also emotional factors. Knowing that fear and anxiety also have an important impact on decision-making, distinct counselling about the procedures, the subsequent risk reduction as well as the psychological effects of prophylactic surgeries are essential.

Breast cancer (BC) patients often ask for a healthy diet. Here, we investigated a healthy standard diet (SD), a low carb diet (LCD), and a ketogenic diet (KD) for BC patients during the rehabilitation phase. KOLIBRI was an open-label non-randomized one-site nutritional intervention trial, combining inpatient and outpatient phases for 20 weeks. Female BC patients (n = 152, mean age 51.7 years) could select their diet. Data collected were: Quality of life (QoL), spiroergometry, body composition, and blood parameters. In total 30, 92, and 30 patients started the KD, LCD, and SD, respectively. Of those, 20, 76, and 25 completed the final examination. Patients rated all diets as feasible in daily life. All groups enhanced QoL, body composition, and physical performance. LCD participants showed the most impressive improvement in QoL aspects. KD participants finished with a very good physical performance and muscle/fat ratio. Despite increased cholesterol levels, KD patients had the best triglyceride/high-density lipoprotein (HDL) ratio and homeostatic model assessment of insulin resistance index (HOMA-IR). Most metabolic parameters significantly improved in the LCD group. SD participants ended with remarkably low cholesterol levels but did not improve triglyceride/HDL or HOMA-IR. In conclusion, both well-defined KDs and LCDs are safe and beneficial for BC patients and can be recommended during the rehabilitation phase.

Background Obesity and low muscle mass are associated with worse outcomes of breast cancer patients. We conducted a controlled trial to study the impact of a ketogenic diet (KD) based on natural foods versus an unspecified standard diet (SD) on body composition in breast cancer patients undergoing radiotherapy. Methods Patients with non-metastasized breast cancer were allocated to either the KD (N = 32) or the SD (N = 31) during radiotherapy. Body composition was measured weekly by bioimpedance analysis. Blood parameters and quality of life were assessed before, during, and at the end of radiotherapy. Results A total of 29 KD and 30 SD patients completed the study. During radiotherapy, mean and median fasting BHB concentrations in the KD group were 0.72 and 0.49 mmol/l (range 0.06–4.9) which was significantly higher than those in the SD group (p −16). There was a very small and insignificant increase in body weight and fat mass in the SD group, as well as a decrease of fat free mass. In contrast, patients in the KD group lost body weight and fat free and skeletal muscle mass quickly after diet onset, which for the most part was related to water losses. The KD did not cause further substantial changes in fat free or skeletal muscle mass, but was associated with a gradual decrease of 0.4 kg body weight and fat mass per week (p p = 6.3 × 10−5). Global quality of life remained stable in the SD group but increased in the KD group from a score of 66.7 to 75.0 (p = 0.20). Conclusions In breast cancer patients undergoing curative radiotherapy, a KD based on natural foods is feasible. After initial water losses, the KD tends to reduce body weight and fat mass while preserving fat free and skeletal muscle mass. Trial registration ClinicalTrials.gov identifier: NCT02516501, registered on August 06, 2015.

Background: Breast Cancer (BC) patients often ask for a healthy diet. Here, we compared three different diets which could support BC patients during the rehabilitation process: a healthy standard diet (SD), a low carb diet (LCD) and a ketogenic diet (KD).Patients and Methods:KOLIBRI was a one-site nutritional intervention trial, combining inpatient and outhouse phases for 20 weeks. Female BC patients (n=152; mean age 51.7 years) could select their diet. Data collected were: quality of life (QoL), spiroergometry, body composition, blood biochemistry. 30, 92 and 30 patients started the KD, LCD and SD, respectively. Of those, 20, 72 and 25 completed the final examination.Results:Patients rated all diets as feasible in daily life. No adverse effects occurred in any diet group. KD offered the highest amount of energy and protein. All groups improved in QoL, body composition and physical performance. KD was superior in improving fatigue, insomnia and physical functioning. KD participants finished with the best physical performance and the highest muscle/fat ratio. Despite their increased cholesterol, KD patients had the best triglyceride/HDL ratio and HOMA-IR. Most metabolic parameters significantly improved in the LCD group. SD participants ended with remarkably low cholesterol levels, but did not improve triglyceride/HDL or HOMA-IR.Conclusions:A well-defined KD and LCD are safe and beneficial for BC. Both diets could thus be recommended for patients wanting to self-support their rehabilitation process.Trial registration IDNCT02092753 Protocol Registration date:02/26/2014Ketogenic Or LOGI Diet In a Breast Cancer Rehabilitation Intervention (KOLIBRI)URL: https://clinicaltrials.gov/ct2/show/NCT02092753?cond=KOLIBRI&draw=2&rank=1

Objective: In proliferative retinopathies, complications derived from neovascularization cause blindness. During early disease, pericyte’s apoptosis contributes to endothelial dysfunction and leakage. Hypoxia then drives VEGF (vascular endothelial growth factor) secretion and pathological neoangiogenesis. Cardiac ANP (atrial natriuretic peptide) contributes to systemic microcirculatory homeostasis. ANP is also formed in the retina, with unclear functions. Here, we characterized whether endogenously formed ANP regulates retinal (neo)angiogenesis. Approach and Results: Retinal vascular development and ischemia-driven neovascularization were studied in mice with global deletion of GC-A (guanylyl cyclase-A), the cGMP (cyclic guanosine monophosphate)-forming ANP receptor. Mice with a floxed GC-A gene were interbred with Tie2-Cre, GFAP-Cre, or PDGF-Rβ-Cre ERT2 lines to dissect the endothelial, astrocyte versus pericyte-mediated actions of ANP in vivo. In neonates with global GC-A deletion (KO), vascular development was mildly delayed. Moreover, such KO mice showed augmented vascular regression and exacerbated ischemia-driven neovascularization in the model of oxygen-induced retinopathy. Notably, absence of GC-A in endothelial cells did not impact retinal vascular development or pathological neovascularization. In vitro ANP/GC-A/cGMP signaling, via activation of cGMP-dependent protein kinase I, inhibited hypoxia-driven astrocyte’s VEGF secretion and TGF-β (transforming growth factor beta)–induced pericyte apoptosis. In neonates lacking ANP/GC-A signaling in astrocytes, vascular development and hyperoxia-driven vascular regression were unaltered; ischemia-induced neovascularization was modestly increased. Remarkably, inactivation of GC-A in pericytes retarded physiological retinal vascularization and markedly enhanced cell apoptosis, vascular regression, and subsequent neovascularization in oxygen-induced retinopathy. Conclusions: Protective pericyte effects of the ANP/GC-A/cGMP pathway counterregulate the initiation and progression of experimental proliferative retinopathy. Our observations indicate augmentation of endogenous pericyte ANP signaling as target for treatment of retinopathies associated with neovascularization.

SummaryDie Ernährung ist ein wichtiger Bestandteil der integrativen Krebstherapie. Doch welche ist die richtige? Sollten Krebspatienten lieber vegetarisch essen oder eher Fleisch? Oder ist viel Fett bei dieser zehrenden Erkrankung das Beste für den Körper? Unsere Ernährungsexperten erklären, welche Vorteile die vegetarische Kost, die Paläo-Diät und die ketogene Diät bei Krebs haben.

SummaryEine ketogene Ernährung mit einem sehr hohen Fettanteil und einer Verknappung von Glukose zeigt tierexperimentell günstige Effekte auf Tumorwachstum und Überleben. In der humanen Krebstherapie deuten sich positive Wirkungen der ketogenen Diät in der Kombination mit Strahlen- und Chemotherapie an.

Introduction: Expression of fibroblast activation protein (FAP) has been detected in activated fibroblasts participating in injury response, fibrotic and inflammatory conditions, and tumorigenesis. Human endometrium is equally characterized by rapid tissue remodeling events due to the reproductive tasks comprising the activity of proteolytic enzymes. Objective: We therefore hypothesized that FAP-positive fibroblasts could also be involved in physiological processes requiring tissue remodeling, such as decidualization during early pregnancy. Methods/Results: The expression of FAP was analyzed by immunohistochemistry in frozen sections of decidual tissue from early pregnancy (gestational weeks: 6–12). All tissue samples clearly displayed a strong expression of FAP on the surface of stromal fibroblasts. Additionally, the percentage of FAP-positive fibroblasts freshly isolated from the decidua of the corresponding gestational weeks was calculated by applying FACS analysis. Decidual fibroblasts of different gestational weeks showed a significant decrease in FAP expression between the 6th and 7th weeks of gestation, which was followed by a steady slow reconstitution. By analyzing the expression of cytokines, chemokines, and growth factors of isolated FAP-positive decidual fibroblasts, we detected high levels of monocyte-attracting chemokines (growth-related oncogene alpha and monocyte chemoattractant protein-1 and -2), granulocyte-attracting chemokines (e.g., IL-8), proinflammatory factors (IL-1α and tumor necrosis factor alpha), and angiogenic substances (e.g., vascular endothelial growth factor and IL-8), which all promote an optimal microenvironment for implantation and growth of the conceptus. Conclusions: Our data demonstrate that the healthy early pregnancy decidua is characterized by a general occurrence of FAP-positive fibroblasts possibly participating in active tissue remodeling during implantation.

Problem Human pregnancy needs a remarkable local immune tolerance toward the conceptus. Myeloid-derived suppressor cells (MDSC) are important players promoting cancer initiation and progression by suppressing T-cell functions and thus inducing immune tolerance. Therefore, MDSC were expected within decidua. Methods Subpopulations of CD33(+) immune cells were isolated from human early pregnancy decidua and characterized phenotypically and functionally by microscopy, FACS analysis, RT-PCR, Western blotting and in the coculture with T cells. Results Decidua harbors CD33(+) /HLA-DR(neg) and CD33(+) /HLA-DR(+/-) cells which both express arginase, iNOS and IDO and a typical cytokine profile. Both subtypes potently suppress T-cell proliferation and therefore fulfill the criteria of MDSC. Conclusion We characterized a new population of CD33(+) /HLA-DR(neg) and CD33(+) /HLA-DR(+/-) cells in human early pregnancy decidua with properties of classical MDSC and thus potentially being an important player in immune tolerance in pregnancy.