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The introduction of robotically assisted surgery was a milestone for minimally invasive surgery in the 21st century. Currently, there are two CE-approved robotically assisted surgery systems for use and development in pediatrics. Specifically, tremor filtration and optimal visualization are approaches which can have enormous benefits for procedures in small bodies. Robotically assisted surgery in children might have advantages compared to laparoscopic or open approaches. This review focuses on the research literature regarding robotically assisted surgery that has been published within the past decade. A literature search was conducted to identify studies comparing robotically assisted surgery with laparoscopic and open approaches. While reported applications in urology were the most cited, three other fields (gynecology, general surgery, and “others”) were also identified. In total, 36 of the publications reviewed suggested that robotically assisted surgery was a good alternative for pediatric procedures. After several years of experience of this surgery, a strong learning curve was evident in the literature. However, some authors have highlighted limitations, such as high cost and a limited spectrum of small-sized instruments. The recent introduction of reusable 3 mm instruments to the market might help to overcome these limitations. In the future, it can be anticipated that there will be a broader range of applications for robotically assisted surgery in selected pediatric surgeries, especially as surgical skills continue to improve and further system innovations emerge.
Bayesian inference is ubiquitous in science and widely used in biomedical research such as cell sorting or “omics” approaches, as well as in machine learning (ML), artificial neural networks, and “big data” applications. However, the calculation is not robust in regions of low evidence. In cases where one group has a lower mean but a higher variance than another group, new cases with larger values are implausibly assigned to the group with typically smaller values. An approach for a robust extension of Bayesian inference is proposed that proceeds in two main steps starting from the Bayesian posterior probabilities. First, cases with low evidence are labeled as “uncertain” class membership. The boundary for low probabilities of class assignment (threshold 𝜀
) is calculated using a computed ABC analysis as a data-based technique for item categorization. This leaves a number of cases with uncertain classification (p < 𝜀
). Second, cases with uncertain class membership are relabeled based on the distance to neighboring classified cases based on Voronoi cells. The approach is demonstrated on biomedical data typically analyzed with Bayesian statistics, such as flow cytometric data sets or biomarkers used in medical diagnostics, where it increased the class assignment accuracy by 1–10% depending on the data set. The proposed extension of the Bayesian inference of class membership can be used to obtain robust and plausible class assignments even for data at the extremes of the distribution and/or for which evidence is weak.
Queck, Alexander David Roger ; Uschner, Frank Erhard ; Ferstl, Philip ; Schulz, Martin ; Brol, Maximilian ; Praktiknjo, Michael ; Schierwagen, Robert ; Klein, Sabine ; Straßburg, Christian P. ; Meyer, Carsten ; Jansen, Christian ; Berres, Marie-Luise ; Trebicka, Jonel
Background: Pathogenesis of portal hypertension is multifactorial and includes pathologic intrahepatic angiogenesis, whereby TIPS insertion is an effective therapy of portal hypertension associated complications. While angiogenin is a potent contributor to angiogenesis in general, little is known about its impact on TIPS function over time. Methods: In a total of 118 samples from 47 patients, angiogenin concentrations were measured in portal and inferior caval vein plasma at TIPS insertion (each blood compartment n = 23) or angiographic intervention after TIPS (each blood compartment n = 36) and its relationship with patient outcome was investigated. Results: Angiogenin levels in the inferior caval vein were significantly higher compared to the portal vein (P = 0.048). Ten to 14 days after TIPS, inferior caval vein angiogenin level correlated inversely with the portal systemic pressure gradient (P<0.001), measured invasively during control angiography. Moreover, patients with TIPS revision during this angiography, showed significantly lower angiogenin level in the inferior caval vein compared to patients without TIPS dysfunction (P = 0.01). Conclusion: In cirrhosis patients with complications of severe portal hypertension, circulating levels of angiogenin are derived from the injured liver. Moreover, angiogenin levels in the inferior caval vein after TIPS may predict TIPS dysfunction.
Over the last decade, cases of metabolic syndrome and type II diabetes have increased exponentially. Exercise and ω-3 polyunsaturated fatty acid (PUFA)-enriched diets are usually prescribed but no therapy is effectively able to restore the impaired glucose metabolism, hypertension, and atherogenic dyslipidemia encountered by diabetic patients. PUFAs are metabolized by different enzymes into bioactive metabolites with anti- or pro-inflammatory activity. One important class of PUFA metabolizing enzymes are the cytochrome P450 (CYP) enzymes that can generate a series of bioactive products, many of which have been attributed protective/anti-inflammatory and insulin-sensitizing effects in animal models. PUFA epoxides are, however, further metabolized by the soluble epoxide hydrolase (sEH) to fatty acid diols. The biological actions of the latter are less well understood but while low concentrations may be biologically important, higher concentrations of diols derived from linoleic acid and docosahexaenoic acid have been linked with inflammation. One potential application for sEH inhibitors is in the treatment of diabetic retinopathy where sEH expression and activity is elevated as are levels of a diol of docosahexaenoic acid that can induce the destabilization of the retina vasculature.
Reggio, Alessio ; Buonomo, Viviana ; Berkane, Rayene ; Bhaskara, Ramachandra M. ; Tellechea, Mariana ; Peluso, Ivana ; Polishchuk, Elena ; Di Lorenzo, Giorgia ; Cirillo, Carmine ; Esposito, Marianna ; Hussain, Adeela ; Hübner, Antje-Kathrin ; Hübner, Christian ; Settembre, Carmine ; Hummer, Gerhard ; Grumati, Paolo ; Stolz, Alexandra
Degradation of the endoplasmic reticulum (ER) via selective autophagy (ER-phagy) is vital for cellular homeostasis. We identify FAM134A/RETREG2 and FAM134C/RETREG3 as ER-phagy receptors, which predominantly exist in an inactive state under basal conditions. Upon autophagy induction and ER stress signal, they can induce significant ER fragmentation and subsequent lysosomal degradation. FAM134A, FAM134B/RETREG1, and FAM134C are essential for maintaining ER morphology in a LC3-interacting region (LIR)-dependent manner. Overexpression of any FAM134 paralogue has the capacity to significantly augment the general ER-phagy flux upon starvation or ER-stress. Global proteomic analysis of FAM134 overexpressing and knockout cell lines reveals several protein clusters that are distinctly regulated by each of the FAM134 paralogues as well as a cluster of commonly regulated ER-resident proteins. Utilizing pro-Collagen I, as a shared ER-phagy substrate, we observe that FAM134A acts in a LIR-independent manner and compensates for the loss of FAM134B and FAM134C, respectively. FAM134C instead is unable to compensate for the loss of its paralogues. Taken together, our data show that FAM134 paralogues contribute to common and unique ER-phagy pathways.
Panuccio, Giuseppe ; De Rosa, Salvatore ; Landmesser, Ulf ; Leistner, David M. ; Abdelwahed, Youssef S.
An 80-year-old post–coronary artery bypass graft (CABG) patient had an acute coronary syndrome with non–ST-segment elevation myocardial infarction (ACS-NSTE) with saphenous vein graft (SVG)–obtuse marginal stenosis. High-definition intravascular ultrasound revealed an underexpanded SVG stent with a hyperechoic structure. Optical coherence tomography confirmed surgical clip causing compression, resolved by post-dilation. This case underscores ACS-NSTE complexity post-CABG and the critical role of coronary imaging in optimizing interventions by addressing surgical clip–induced compression.
Oberoi-Khanuja, Tripat Kaur ; Karreman, Christiaan ; Larisch, Sarit ; Rapp, Ulf Rüdiger ; Rajalingam, Krishnaraj
Inhibitor of apoptosis (IAPs) proteins are characterized by the presence of evolutionarily conserved baculoviral inhibitor of apoptosis repeat (BIR) domains, predominantly known for their role in inhibiting caspases and, thereby, apoptosis. We have shown previously that multi-BIR domain-containing IAPs, cellular IAPs, and X-linked IAP can control tumor cell migration by directly regulating the protein stability of C-RAF kinase. Here, we extend our observations to a single BIR domain containing IAP family member melanoma-IAP (ML-IAP). We show that ML-IAP can directly bind to C-RAF and that ML-IAP depletion leads to an increase in C-RAF protein levels, MAPK activation, and cell migration in melanoma cells. Thus, our results unveil a thus far unknown role for ML-IAP in controlling C-RAF stability and cell migration.
Cancer microenvironment is now recognized as a critical regulator of all stages of cancer development. Beside the tumor vasculature and tumor-infiltrating immune cells, other stromal cells such as cancer-associated fibroblasts (CAFs) regulate tumor growth. Fibroblasts are ubiquitous cells in connective tissue, where they shape the extracellular matrix (ECM). Fibroblasts are usually quiescent but get activated when tissue homeostasis is disturbed. Then, activated fibroblasts rebuild the ECM and communicate with local cells to participate in wound repair. These repair properties can go awry when being unchecked, which can lead to fibrosis and subsequently cancer development. CAFs can promote cancer development by fostering tumor cell growth, polarizing immune cells to an immunosuppressive phenotype, and crosslinking collagen to enable tumor cell invasion. Molecular mechanisms of CAF activation, thus, need to be understood to target these cells in tumors. Prostanoid prostaglandin E2 (PGE2) is viewed as a pro-tumor lipid mediator as suggested by studies pharmacologically or genetically targeting the enzymes producing PGE2, such as microsomal PGE synthase-1 (mPGES-1) in tumor models. Similar to CAFs, PGE2 drives tumor cell growth and tumor-associated immune suppression. Therefore, I hypothesized that PGE2 may play a role in CAF activation.
This hypothesis was tested in two mouse models of breast cancer (orthotopic grafting model, and polyoma middle T oncogene transgenic model), besides using isolated mammary gland (MG) fibroblasts in vitro. As expected, given the pro-tumor function of PGE2, knocking out mPGES-1 reduced the growth of oncogene-driven and transplanted mammary tumors. Surprisingly, CAF density was markedly increased when mPGES-1 was depleted. Importantly, despite reduced primary tumor growth, I observed enhanced lung metastasis upon mPGES-1depletion. Using MG-derived fibroblasts in vitro furthermore revealed that treatment with PGE2 reduced a TGFβtriggered CAF-like activation state. Importantly, bioinformatics analysis of a human breast cancer patient dataset revealed a negative correlation of a PGE2 production signature with fibroblast marker genes. In a next step I investigated if the increased CAF infiltrate was connected to the reduced tumor growth upon depletion of PGE2. To unravel this, I first asked through which E prostanoid (EP) receptor PGE2 signals in fibroblasts. MG fibroblasts mainly expressed EP3, and EP3 KO fibroblasts showed a hyper-proliferative and activated phenotype, indicating EP3 as the main PGE2 receptor in MG fibroblasts. Co-injecting of EP3 KO MG fibroblasts and tumor cells in WT mice suppressed tumor growth, whereas co-injection of WT fibroblasts with tumor cell in mPGES-1 KO mice increased tumor growth. These data indicate that PGE2 restricts CAF levels through EP3, which supports tumor growth. Whole transcriptome mRNAsequencing of WT and mPGES-1 KO FACS-sorted CAFs combined with immunohistochemical data suggested a role of p38 mitogen-activated protein kinase (MAPK) in the modulation of fibroblast activation by PGE2.
In summary, I showed in two breast cancer models that mPGES-1 depletion delays breast cancer progression, which is probably driven by the EP3-PGE2 signaling axis in host stroma. PGE2 appears to be a potent anti-fibroblast activation agent in tumors via EP3 and downstream p38 MAPK signaling. This study therefore hits the dogmatic perception of the general pro-tumor nature of PGE2; showing that PGE2 might be a double-edged mediator that can promote tumor growth at the primary site by restricting CAF expansion, which may in turn hinder infiltration of tumor cells to a secondary site.
Raimann, Florian ; Lindner, Gesche Marie-Louise ; Martin, Christoph ; Jennewein, Lukas ; Lustenberger, Thomas ; Piekarski, Florian ; Zacharowski, Kai ; Weber, Christian Friedrich
Background: Acute bleeding requires fast and targeted therapy. Therefore, knowledge of the patient's potential to form a clot is crucial. Point-of-care testing (POCT) provides fast and reliable information on coagulation. Structural circumstances, such as person-bound sample transport, can prolong the reporting of the results. The aim of the present study was to investigate the diagnostic quality and accuracy between POCT INR diagnostics and standard laboratory analysis (SLA) as well as the time advantage between a pneumatic tube and a personal-based transport system. Methods: Two groups of haemorrhagic patients (EG: emergency department; OG: delivery room; each n = 12) were examined in the context of bleeding emergencies using POCT and SLA. Samples were transported via a pneumatic tube system or by a personal transport service. Results: INR results between POCT and SLA showed a high and significant correlation (EG: p < 0.001; OG: p < 0.001). POCT results were reported significantly more quickly (EG: 1.1 vs. 39.6 min; OG: 2.0 vs. 75.0 min; p < 0.001) and required less time for analysis (EG: 0.3 vs. 24.0 min; OG: 0.5 vs. 45.0 min; p < 0.001) compared to SLA. The time for transportation with the pneumatic tube was significantly shorter (8.0 vs. 18.5 min; p < 0.001) than with the personal-based transport system. Conclusion: The results of the present study suggest that POCT may be a suitable method for the emergency diagnosis and may be used as prognostic diagnostic elements in haemotherapy algorithms to initiate targeted haemotherapy at an early point in time.
Classical molecular dynamics (MD) simulations provide unmatched spatial and time resolution of protein structure and function. However, accuracy of MD simulations often depends on the quality of force field parameters and the time scale of sampling. Another limitation of conventional MD simulations is that the protonation states of titratable amino acid residues remain fixed during simulations, even though protonation state changes coupled to conformational dynamics are central to protein function. Due to the uncertainty in selecting protonation states, classical MD simulations are sometimes performed with all amino acids modeled in their standard charged states at pH 7. Here we performed and analyzed classical MD simulations on high-resolution cryo-EM structures of two membrane proteins that transfer protons by catalyzing protonation/deprotonation reactions. In simulations performed with amino acids modeled in their standard protonation state the structure diverges far from its starting conformation. In comparison, MD simulations performed with pre-determined protonation states of amino acid residues reproduce the structural conformation, protein hydration, and protein-water and protein-protein interactions of the structure much better. The results suggest it is crucial to perform basic protonation state calculations, especially on structures where protonation changes play an important functional role, prior to launching any MD simulations. Furthermore, the combined approach of protonation state prediction and MD simulations can provide valuable information on the charge states of amino acids in the cryo-EM sample. Even though accurate prediction of protonation states currently remains a challenge, we introduce an approach of combining pKa prediction with cryo-EM density map analysis that helps in improving not only the protonation state predictions, but also the atomic modeling of density data.