Tofacitinib Affects M1-like and M2-like Polarization and Tissue Factor Expression in Macrophages of Healthy Donors and IBD Patients.

BACKGROUND: Tofacitinib, as inhibitor of Janus kinases (JAK), interrupts the transmission of numerous pro-inflammatory cytokines involved in the pathogenesis of inflammatory bowel diseases (IBD). Therefore, tofacitinib provides a potent option to treat ulcerative colitis (UC). Besides the anti-inflammatory potential, inhibition of widespread JAKs carries the risk of side effects. Macrophages are involved in the form of different subtypes in inflammation, wound healing, and even coagulation. This study aimed to explore the balanced use of tofacitinib in M1-like as well as M2-like macrophages of healthy donors and patients with IBD. METHODS: Monocytes of healthy donors and patients with chronic courses of IBD were obtained from blood samples. Macrophage colony-stimulating factor (M-CSF)-derived macrophages were treated with tofacitinib (1 µM, 5 µM, 10 µM) and polarized with either lipopolysaccharide and interferon (IFN)-γ towards M1-like-phenotype or with interleukin (IL)-4 towards M2-like-phenotype. ELISA and flow cytometry were used to evaluate cytokine levels and surface molecules. RESULTS: Tofacitinib had a modulating effect on M1-like macrophages whereby the effect on pro-inflammatory cytokines (TNF-α, IL-6, IL-1ß, IL-12, IL-23) was less pronounced than the induction of anti-inflammatory IL-10. However, during M2-like polarization tofacitinib impaired the development of the corresponding phenotype becoming evident through decreased IL-10 levels and CD206 expression in treated macrophages. In both phenotypes, tofacitinib strongly downregulated the expression of immunostimulatory molecules (CD80, CD86, CD83, CD40). Furthermore, a dose-dependent correlation between treatment with tofacitinib and expressed tissue factor was noticed. CONCLUSIONS: Tofacitinib influences both polarizations (M1/M2) and the expression of tissue factor in a dose-dependent manner.

This study revealed a dose-dependent effect of tofacitinib on both M1-like and M2-like polarization, resulting in a decreased development of the corresponding phenotype. Furthermore, the applied dose of tofacitinib correlated with the expressed tissue factor in M1-like macrophages.

Anatomical principles of ankle denervation - An update.

BACKGROUND: Denervation is a surgical option in ankle arthrosis when conservative therapy has failed. Sectioning all joint branches is essential for its success. The locations of the articular branches of the saphenous (Sa), tibial (Ti), sural (Su), superficial (Ps) and deep peroneal (Pp) nerves are specified. METHODS: In 16 cryopreserved specimens, the courses of the nerves were prepared. Their articular branches were identified, and their respective locations documented by using a new reference system. RESULTS: The articular branches to the ankle ranged from 5 to 30 cm measured from the foot sole. The Sa should be transected at 22.5 cm, the Su at 20 cm, and the Pp at 15 cm. The Ti should be skeletonized up to 25 cm. Epifascial dissection of the Ps is to be performed below 15 cm. CONCLUSION: The study specifies the joint branches of the ankle in an intraoperatively reproducible reference system and thus minimizes the required skin incisions.

A pharmacokinetic-pharmacodynamic model for chemoprotective agents against malaria.

Chemoprophylactics are a vital tool in the fight against malaria. They can be used to protect populations at risk, such as children younger than the age of 5 in areas of seasonal malaria transmission or pregnant women. Currently approved chemoprophylactics all present challenges. There are either concerns about unacceptable adverse effects such as neuropsychiatric sequalae (mefloquine), risks of hemolysis in patients with G6PD deficiency (8-aminoquinolines such as tafenoquine), or cost and daily dosing (atovaquone-proguanil). Therefore, there is a need to develop new chemoprophylactic agents to provide more affordable therapies with better compliance through improving properties such as pharmacokinetics to allow weekly, preferably monthly, dosing. Here we present a pharmacokinetic-pharmacodynamic (PKPD) model constructed using DSM265 (a dihydroorotate dehydrogenase inhibitor with activity against the liver schizonts of malaria, therefore, a prophylaxis candidate). The PKPD model mimics the parasite lifecycle by describing parasite dynamics and drug activity during the liver and blood stages. A major challenge is the estimation of model parameters, as only blood-stage parasites can be observed once they have reached a threshold. By combining qualitative and quantitative knowledge about the parasite from various sources, it has been shown that it is possible to infer information about liver-stage growth and its initial infection level. Furthermore, by integrating clinical data, the killing effect of the drug on liver- and blood-stage parasites can be included in the PKPD model, and a clinical outcome can be predicted. Despite multiple challenges, the presented model has the potential to help translation from preclinical to late development for new chemoprophylactic candidates.

Surface Design Options in Polymer- and Lipid-Based siRNA Nanoparticles Using Antibodies.

The mechanism of RNA interference (RNAi) could represent a breakthrough in the therapy of all diseases that arise from a gene defect or require the inhibition of a specific gene expression. In particular, small interfering RNA (siRNA) offers an attractive opportunity to achieve a new milestone in the therapy of human diseases. The limitations of siRNA, such as poor stability, inefficient cell uptake, and undesired immune activation, as well as the inability to specifically reach the target tissue in the body, can be overcome by further developments in the field of nanoparticulate drug delivery. Therefore, types of surface modified siRNA nanoparticles are presented and illustrate how a more efficient and safer distribution of siRNA at the target site is possible by modifying the surface properties of nanoparticles with antibodies. However, the development of such efficient and safe delivery strategies is currently still a major challenge. In consideration of that, this review article aims to demonstrate the function and targeted delivery of siRNA nanoparticles, focusing on the surface modification via antibodies, various lipid- and polymer-components, and the therapeutic effects of these delivery systems.

Parasites of pufferfish, Lagocephalus spp. and Torquigener flavimaculosus of the Israeli Mediterranean: A new case of Lessepsian endoparasites.

With the opening of the Suez Canal as a link between the Red Sea and the Mediterranean Sea in 1869, the biogeographical event of the Lessepsian migration has been starting. Aided by beneficial conditions in the new habitat, almost 500 marine species have immigrated and often established themselves in the Mediterranean Sea, including several pufferfish species, with all of them extending their range and becoming important components of the local fauna. The parasitic fauna of these pufferfish has scarcely been examined in the Mediterranean Sea or in their native range, which provides the opportunity to study host-parasite interaction in a new habitat. The present study describes the parasitic fauna in four alien invasive pufferfish species (Lagocephalus guentheri, L. sceleratus, L. suezensis, and Torquigener flavimaculosus) of various sizes and ages on the Israeli Mediterranean coast. The parasite fauna of these species was diverse (Maculifer dayawanensis Digenea; Calliterarhynchus gracilis, Nybelinia africana and Tetraphyllidea larvae Cestoda; Hysterothylacium reliquens, Hysterothylacium sp. and Raphidascaris sp. Nematoda; Trachellobdella lubrica Hirudinea and Caligus fugu and Taeniacanthus lagocephali Copepoda) and consisted of mostly generalist species, most likely acquired in the new habitat, and specialist copepod ectoparasites, having co-invaded with the pufferfish. Additionally, the oioxenic opecoelid digenean Maculifer dayawanensis was found in two pufferfish species. The genus was previously only known from the Indo-Pacific Ocean, representing the eighth reported case of a Lessepsian endoparasite so far. Our results suggest a change in parasite fauna to native Mediterranean species in the pufferfish like previously reported in other Lessepsian migrant predatory fish species and a wider spread of co-invasion of fish endoparasites to the Mediterranean Sea than previously assumed. The study also provides several new host records and the first report for parasites in T. flavimaculosus.

The Memory T Cell "Communication Web" in Context with Gastrointestinal Disorders-How Memory T Cells Affect Their Surroundings and How They Are Influenced by It.

Gut-related diseases like ulcerative colitis, Crohn's disease, or colorectal cancer affect millions of people worldwide. It is an ongoing process finding causes leading to the development and manifestation of those disorders. This is highly relevant since understanding molecular processes and signalling pathways offers new opportunities in finding novel ways to interfere with and apply new pharmaceuticals. Memory T cells (mT cells) and their pro-inflammatory properties have been proven to play an important role in gastrointestinal diseases and are therefore increasingly spotlighted. This review focuses on mT cells and their subsets in the context of disease pathogenesis and maintenance. It illustrates the network of regulatory proteins and metabolites connecting mT cells with other cell types and tissue compartments. Furthermore, the crosstalk with various microbes will be a subject of discussion. Characterizing mT cell interactions will help to further elucidate the sophisticated molecular and cellular networking system in the intestine and may present new ideas for future research approaches to control gut-related diseases.

Nematode Parasites in Baltic Sea Mammals, Grey Seal (Halichoerus grypus (Fabricius, 1791)) and Harbour Porpoise (Phocoena phocoena (L.)), from the German Coast.

PURPOSE: Endoparasitic nematodes of six harbour porpoises Phocoena phocoena and four grey seals Halichoerus grypus, stranded at the eastern coast of the Baltic Sea in Germany in winter 2019, were analysed in order to identify nematode parasites and to compare with recent studies from the same area. METHODS: Endoparasitic nematodes were identified by using both morphological and molecular characters. The successfully obtained sequences of the rDNA marker regions ITS-1, 5.8S, ITS-2 from 29 anisakid and the rDNA marker region ITS-2 of 11 pseudalid nematodes were amplified. RESULTS: Analyses revealed the presence of three parasite species, the anisakid nematode Contracaecum osculatum from grey seals and the pseudalid nematodes Pseudalius inflexus and Stenurus minor from the harbour porpoises. Other anisakid nematodes regularly occurring in the Baltic Sea, e.g. Anisakis simplex or Pseudoterranova decipiens, were not found. CONCLUSIONS: The prevalence of 100% and a severe parasite load in grey seals demonstrated a very high C. osculatum infection of Baltic Sea fish as their regular prey. Prevalence of 33% for parasites in harbour porpoises and minor infection rates, combined with a distinct lack of anisakid nematodes, are typical for the current situation of the porpoise parasite fauna in the Baltic Sea. Invasive parasite species as possible indicators for climate change could not be detected.

Feasibility of CardioSecur®, a Mobile 4-Electrode/22-Lead ECG Device, in the Prehospital Emergency Setting.

Background: This study explores the application of CardioSecur® (CS-ECG), a hand-held 4-electrode/22-lead ECG-device, in comparison with conventional 12-lead electrocardiogram (c12L-ECG) in patients with acute chest pain in the prehospital emergency setting. Methods: CS-ECG systems were provided for two physician-staffed emergency ambulances and parallel recordings of c12L-ECG and CS-ECG were obtained from all patients with acute chest pain. Treating emergency physicians were asked to evaluate the CS-ECG system with a standardized questionnaire. Following study completion, acquired ECGs were analyzed separately by two independent cardiologists blinded to all other medical records. Results: Over a period of 20 months a total of 203 patients were included in our study. According to a standardized questionnaire, 79% of emergency medical professionals preferred application of CS-ECG, with 87% of teams judging CS-ECG to be beneficial for patients. Morover, 79% of physicians reported a reduction in time to definitive diagnosis with implementation of CS-ECG. The majority of professional users attested user-friendliness and feasibility of CS-ECG in terms of easy general handling (94%), application (93%), and placement of electrodes (98%). During prehospital triage, both c12L-ECG and CS-ECG correctly identified 31 (91%) patients with ST-elevation myocardial infarction (STEMI). Conclusion: In this first pilot study, implementation of the CardioSecur®-ECG system in the prehospital emergency setting demonstrated feasibility and user-friendliness so that emergency teams generally preferred CS-ECG to c12L-ECG. Diagnostic yield of CS-ECG was similar to c12L-ECG recordings.

Prediction of mortality rate in acute type A dissection: the German Registry for Acute Type A Aortic Dissection score.

OBJECTIVES: The goal was to develop a scoring system to predict the 30-day mortality rate for patients undergoing surgery for acute type A aortic dissection on the basis of the German Registry for Acute Type A Aortic Dissection (GERAADA) data set and to provide a Web-based application for standard use. METHODS: A total of 2537 patients enrolled in GERAADA who underwent surgery between 2006 and 2015 were analysed. Variable selection was performed using the R-package FAMoS. The robustness of the results was confirmed via the bootstrap procedure. The coefficients of the final model were used to calculate the risk score in a Web-based application. RESULTS: Age [odds ratio (OR) 1.018, 95% confidence interval (CI) 1.009-1.026; P < 0.001; 5-year OR: 1.093], need for catecholamines at referral (OR 1.732, 95% CI 1.340-2.232; P < 0.001), preoperative resuscitation (OR 3.051, 95% CI 2.099-4.441; P < 0.001), need for intubation before surgery (OR 1.949, 95% CI 1.465-2.585; P < 0.001), preoperative hemiparesis (OR 1.442, 95% CI 0.996-2.065; P = 0.049), coronary malperfusion (OR 1.870, 95% CI 1.386-2.509; P < 0.001), visceral malperfusion (OR 1.748, 95% CI 1.198-2.530; P = 0.003), dissection extension to the descending aorta (OR 1.443, 95% CI 1.120-1.864; P = 0.005) and previous cardiac surgery (OR 1.772, 95% CI 1.048-2.903; P = 0.027) were independent predictors of the 30-day mortality rate. The Web application based on the final model can be found at https://www.dgthg.de/de/GERAADA_Score. CONCLUSIONS: The GERAADA score is a simple, effective tool to predict the 30-day mortality rate for patients undergoing surgery for acute type A aortic dissection. We recommend the widespread use of this Web-based application for standard use.

FAMoS: A Flexible and dynamic Algorithm for Model Selection to analyse complex systems dynamics.

Most biological systems are difficult to analyse due to a multitude of interacting components and the concomitant lack of information about the essential dynamics. Finding appropriate models that provide a systematic description of such biological systems and that help to identify their relevant factors and processes can be challenging given the sheer number of possibilities. Model selection algorithms that evaluate the performance of a multitude of different models against experimental data provide a useful tool to identify appropriate model structures. However, many algorithms addressing the analysis of complex dynamical systems, as they are often used in biology, compare a preselected number of models or rely on exhaustive searches of the total model space which might be unfeasible dependent on the number of possibilities. Therefore, we developed an algorithm that is able to perform model selection on complex systems and searches large model spaces in a dynamical way. Our algorithm includes local and newly developed non-local search methods that can prevent the algorithm from ending up in local minima of the model space by accounting for structurally similar processes. We tested and validated the algorithm based on simulated data and showed its flexibility for handling different model structures. We also used the algorithm to analyse experimental data on the cell proliferation dynamics of CD4+ and CD8+ T cells that were cultured under different conditions. Our analyses indicated dynamical changes within the proliferation potential of cells that was reduced within tissue-like 3D ex vivo cultures compared to suspension. Due to the flexibility in handling various model structures, the algorithm is applicable to a large variety of different biological problems and represents a useful tool for the data-oriented evaluation of complex model spaces.

Investigating the Dynamics of MCMV-Specific CD8+ T Cell Responses in Individual Hosts.

Infection by Cytomegalovirus (CMV) is characterized by the massive expansion and continued maintenance of CMV-specific CD8+ T cells for certain CMV-derived peptides. This phenomenon called "memory inflation" has made CMV a primary target for the generation of T cell based vaccine vectors against various diseases. However, many aspects concerning the generation and maintenance of the inflationary CD8+ T cell response still remain to be resolved. In this study, we combined experimental data and mathematical models to analyze the dynamics of circulatory inflationary CD8+ T cells within individual mice infected by MCMV. Obtaining frequent measurements on the number and frequency of CMV-specific CD8+ T cells up to 70 days post infection, we find that mathematical models assuming differing viral stimuli during acute infection and the inflationary phase provide a better description for the observed dynamics than models relying on similar viral stimuli during both phases. In addition, our analysis allowed a detailed quantification of the different phases of memory inflation within individual mice (1st-expansion - contraction - 2nd expansion/maintenance) indicating remarkable consistency of the timing of these phases across mice, but considerable variation in the size of the individual responses between mice. Our analysis provides a first step toward generating a mechanistic framework for analyzing the generation and maintenance of inflationary CD8+ T cells while accounting for individual heterogeneity. Extending these analyses by incorporating measurements from additional compartments and more prolonged sampling will help to obtain a systematic and quantitative understanding of the factors regulating the process of memory inflation.

Varying Immunizations With Plasmodium Radiation-Attenuated Sporozoites Alter Tissue-Specific CD8+ T Cell Dynamics.

Whole sporozoite vaccines represent one of the most promising strategies to induce protection against malaria. However, the development of efficient vaccination protocols still remains a major challenge. To understand how the generation of immunity is affected by variations in vaccination dosage and frequency, we systematically analyzed intrasplenic and intrahepatic CD8+ T cell responses following varied immunizations of mice with radiation-attenuated sporozoites. By combining experimental data and mathematical modeling, our analysis indicates a reversing role of spleen and liver in the generation of protective liver-resident CD8+ T cells during priming and booster injections: While the spleen acts as a critical source compartment during priming, the increase in vaccine-induced hepatic T cell levels is likely due to local reactivation in the liver in response to subsequent booster injections. Higher dosing accelerates the efficient generation of liver-resident CD8+ T cells by especially affecting their local reactivation. In addition, we determine the differentiation and migration pathway from splenic precursors toward hepatic memory cells thereby presenting a mechanistic framework for the impact of various vaccination protocols on these dynamics. Thus, our work provides important insights into organ-specific CD8+ T cell dynamics and their role and interplay in the formation of protective immunity against malaria.

More or less-On the influence of labelling strategies to infer cell population dynamics.

The adoptive transfer of labelled cell populations has been an essential tool to determine and quantify cellular dynamics. The experimental methods to label and track cells over time range from fluorescent dyes over congenic markers towards single-cell labelling techniques, such as genetic barcodes. While these methods have been widely used to quantify cell differentiation and division dynamics, the extent to which the applied labelling strategy actually affects the quantification of the dynamics has not been determined so far. This is especially important in situations where measurements can only be obtained at a single time point, as e.g. due to organ harvest. To this end, we studied the appropriateness of various labelling strategies as characterised by the number of different labels and the initial number of cells per label to quantify cellular dynamics. We simulated adoptive transfer experiments in systems of various complexity that assumed either homoeostatic cellular turnover or cell expansion dynamics involving various steps of cell differentiation and proliferation. Re-sampling cells at a single time point, we determined the ability of different labelling strategies to recover the underlying kinetics. Our results indicate that cell transition and expansion rates are differently affected by experimental shortcomings, such as loss of cells during transfer or sampling, dependent on the labelling strategy used. Furthermore, uniformly distributed labels in the transferred population generally lead to more robust and less biased results than non-equal label sizes. In addition, our analysis indicates that certain labelling approaches incorporate a systematic bias for the identification of complex cell expansion dynamics.

[The retrocapital osteotomy ("chevron") for correction of splayfoot with hallux valgus]. / Die retrokapitale Osteotomie (Chevron) zur Korrektur der Spreizfussdeformität mit Hallux valgus.

OBJECTIVE: Surgical treatment of hallux valgus deformity with a distal osteotomy of the first metatarsal to address an increased intermetatarsal angle (IMA) I-II. This procedure is combined with a soft-tissue procedure at the first metatarsophalangeal joint: realignment of the first ray, lateral displacement of the first metatarsal head above the sesamoids, rebalancing of the soft tissues at the metatarsophalangeal joint. INDICATIONS: Pain and soft-tissue inflammation at the bunion, impaired function of the metatarsophalangeal joint, and lateral deviation of the hallux. IMA I-II 10 degrees. CONTRAINDICATIONS: Symptomatic osteoarthritis of the first metatarsophalangeal joint, assessed clinically or radiographically. Acute inflammation of the forefoot, osteoporosis of the first metatarsal. Vascular disturbance. Cosmetic indication only. Relative: hypermobility of the first ray, valgus malalignment of the hindfoot, previous retrocapital osteotomy. SURGICAL TECHNIQUE: Lateral soft-tissue release. Resection of the medial pseudoexostosis. V-shaped osteotomy of the distal metatarsal I. Exostosectomy. Lateral displacement of the first metatarsal head. Screw fixation. Realignment of the metatarsophalangeal joint by tightening of the medial soft tissues. POSTOPERATIVE MANAGEMENT: Postoperative shoe with full weight bearing. Active exercises of the foot and hallux. Physiotherapy. Prophylaxis of deep vein thrombosis depending on the degree of mobility. Radiographic control after 6 weeks. Bandage or orthosis to maintain toe alignment. RESULTS: IMA I-II was reduced from 13.6 degrees preoperatively to 6.6 degrees postoperatively. HVA decreased from 29.8 degrees to 8.2 degrees postoperatively.

Conjugated linoleic acid isomers differ in their free radical scavenging properties.

Conjugated linoleic acid (CLA) isomers were investigated for free radical scavenging properties against the stable 2,2-diphenyl-1-picryhydrazyl radical (DPPH(*)) by electron spin resonance (ESR) spectrometry and spectrophotometric methods. ESR measurements confirmed that both c9,t11-CLA and t10,c12-CLA directly reacted with and quenched DPPH radicals, whereas spectrophotometric analysis demonstrated that c9,t11-CLA and t10,c12-CLA differed in their kinetic and thermodynamic properties in reacting with DPPH radicals. t10,c12-CLA was shown to exhibit a greater initial velocity in CLA-DPPH radical reactions at levels of 2.5-80 mg/mL, and c9,t11-CLA scavenged more DPPH radicals at steady state. Similar dose and time relationships were observed for both isomers. In addition, a mixture of c9,t11- and t10,c12-CLA isomers demonstrated a greater initial velocity in quenching DPPH radicals than either isomer alone on the same concentration basis, suggesting that a synergistic effect between CLA isomers existed in their reactions with DPPH radicals. These results support the conclusion that individual CLA isomers differ in their biological actions and indicate that interaction(s) between isomers may contribute to their beneficial effects.