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Unlike the native surface of the implant material (Ti6Al4V), oxidation with H2O2 leads to increased binding of the effective antimicrobial agent poly(hexamethylene) biguanide [PHMB]. However, treating with NaOH instead results in an even higher PHMB mass coverage. After oxidation with H2O2, strong differences in the PHMB adsorption capability between polished and corundum-blasted surfaces appear, indicating a roughness dependence. After NaOH treatment, no such effect was observed. The wetting properties of specimens treated with either H2O2 or NaOH prior to PHMB exposure clearly varied. To unravel the nature of this interaction, widespread in silico and in vitro experiments were performed. Methods: By X-ray photoelectron spectroscopy, scanning electron microscopy, water contact angle measurements and MD simulations, we characterized the interplay between the polycationic antimicrobial agent and the implant surface. A theoretical model for PHMB micelles is tested for its wetting properties and compared to carbon contaminated TiO2. In addition, quantitation of anionic functional group equivalents, the binding properties of PHMB with blocked amino end-group, and the ability to bind chlorhexidine digluconate (CHG) were investigated. Ultimately, the capability of osteoblasts to build calcium apatite, and the activity of alkaline phosphatase on PHMB coated specimens, were determined. Results: Simulated water contact angles on carbon contaminated TiO2 surfaces and PHMB micelle models reveal little influence of PHMB on the wetting properties and point out the major influence of remaining and recovering contamination from ambient air. Testing PHMB adsorption beyond the critical micelle concentration and subsequent staining reveals an island-like pattern with H2O2 as compared to an evenly modified surface with NaOH. Both CHG and PHMB, with blocked amino end groups, were adsorbed on the treated surfaces, thus negating the significant influence of PHMB’s terminal groups. The ability of osteoblasts to produce calcium apatite and alkaline phosphatase is not negatively impaired for PHMB mass coverages up to 8 μg/specimen. Conclusion: Differences in PHMB adsorption are triggered by the number of anionic groups and carbon contaminants, both of which depend on the specimen pre-treatment. With more PHMB covering, the implant surface is protected against the capture of new contamination from the ambient air, thus building a robust antimicrobial and biocompatible surface coating.
Purpose: To (1) describe the prevalence of abnormal sleep quality in patients with hip abductor tears (HAT), to (2) determine whether sleep quality improves after open HAT repair, and to (3) to report clinical short-term outcomes in patients undergoing open HAT repair. Methods: The data of 28 patients (29 hips) who underwant open HAT repair were prospectively analyzed at midterm follow-up. The Pittsburgh Sleep Quality Index (PSQI), modified Harris Hip Score (mHHS), the University of California, Los Angeles activity scale (UCLA), and Visual Analog Scale (VAS) for pain were determined via questionnaire. Paired t-tests were applied to compare preoperative and post-operative Patient-reported Outcome Measures (PROMs). Logistic regression was performed to determine the association between PSQI improvement achievement and demographic variables (laterality, sex, age, body-mass-index (BMI), and preoperative mHHS). The minimal clinically important difference (MCID) was calculated for the mHHS. Results: A total of 28 patients were included. Four patients (14.3%) suffered post-operative complications after open HAT repair. The predominance of patients was female (77.4%), with a mean age of 60 ± 13 years. The average follow-up was 30.35 ± 16.62 months. Preoperatively, 27 (96.4%) patients experienced poor sleep quality (PSQI > 5); at follow-up, 7 (25%) patients experienced poor sleep quality. Univariate logistical regression analysis demonstrated no significant association between preoperative demographic data and achieving postoperative PSQI < 5. The MCID of mHHS was calculated to be 12.5. Overall, 90% of patients achieved MCID for mHHS. Conclusion: Preoperative sleep quality was impaired in 96.4% of HAT patients (PSQI > 5). However, these patients showed an improvement in sleep disturbances after open HAT repair in the early postoperative period. Ninety percent of patients showed significant improvements in mHHS and achieved the corresponding MCID. Level of Evidence: Case series; Level IV.
Background:
Arthroscopic treatment of femoroacetabular impingement syndrome (FAIS) has become a common procedure. However, meaningful long-term clinical outcomes have not been defined.
Purpose:
To define the minimal clinically important difference (MCID), substantial clinical benefit (SCB), and patient acceptable symptomatic state (PASS) for the modified Harris Hip Score (mHHS) at a minimum 10-year follow-up in patients undergoing arthroscopic treatment for FAIS and identify preoperative predictors for achievement of the MCID, SCB, and PASS.
Study Design:
Case-control study; Level of evidence, 3.
Methods:
A consecutive series of patients undergoing arthroscopic treatment for FAIS between 2007 and 2009 with a minimum 10-year follow-up was analyzed. Patient data included patient characteristics, radiographic parameters, and the pre- and postoperative mHHS and visual analog scale (VAS) for pain score. Paired t tests were used to compare the patient-reported outcome measures (PROMs). The MCID was determined by calculating half of the standard deviation, and SCB and PASS were calculated by the anchor method. Correlation and logistic regression analyses were conducted to identify predictors for the achievement of the MCID, SCB, and PASS.
Results:
A total of 44 patients (27 men, 17 women) were included. The mean age and body mass index were 42.2 years (range, 16-67 years) and 22.3 kg/m2 (range, 16.76-29.78 kg/m2), respectively. The MCID, absolute SCB, net change SCB, and PASS of the mHHS were calculated to be 19.6, 90.1, 31.5, and 84.4 points, respectively. Preoperative symptom duration was identified as an independent predictor for the achievement of meaningful clinical outcomes. The median symptom durations for patients who achieved the MCID, absolute SCB, net change SCB, and PASS were 11.7, 9.1, 9.0, and 10.8 months, respectively. The median symptom duration for patients who did not achieve the MCID, absolute SCB, net change SCB, and PASS were 15.8, 17.4, 17.3, and 18.4 months, respectively. No other statistically significant correlations were found.
Conclusion:
The preoperative duration of symptoms was identified as an independent predictor for achievement of the MCID, SCB, and PASS. These findings can be helpful in accelerating the transition to surgical treatment of FAIS.
Adaptation mechanisms within the B cell composition for successful human and murine pregnancies.
(2021)
Introduction
A well-balanced immune maternal status is essential for favourable outcome of pregnancy. Due to their complexities, not all immune adaptations that promote tolerance during pregnancy are known. To understand the adaptation of the B cell compartment, we analysed and compared B cell lymphopoiesis in different lymphoid tissues in a number of murine models.
Furthermore, we focused on the humoral immune response during pregnancy. We analysed immunoglobulin profiles in human subjects and mice during pregnancy.
These cellular alterations are subject to the influence of chemokines, among others. Therefore, we assessed serum levels of B cell activation factor to clarify its effects during pregnancy.
Methods
For analysis of the human peripheral B cell compartment, peripheral blood samples from age-matched non-pregnant and pregnant women without pregnancy complications, immunological disease or acute/chronic inflammation were collected and sub-classified into four different groups: non-pregnant, and first, second, or third trimester of pregnancy. The experiments, based on a mouse model, were performed with 8-week-old female mice: clinically healthy non-pregnant (CBA/J (H2k)), pregnant mice with normal gestation (BALB/c (H2d) x CBA/J (H2k)), and mice with pregnancy loss (DBA/2J (H2d) x CBA/J (H2k)). Subsequently, peripheral blood mononuclear cells from blood and lymphatic organs were isolated following standard protocols. The B cell analysis was performed by flow cytometry. The immunoglobulin serum levels of the human and murine subgroups were quantitated using Bio-Plex isotyping assay and analysed by a Bio-Plex reader. To quantify B cell activating factor (BAFF) in serum of pregnant and non-pregnant mice a BAFF enzyme-linked immunosorbent assay was used. The concentrations were determined by using a FLUOstar OPTIMA microplate reader. All statistical analyses were performed using the Kruskal–Wallis test with Dunn’s post-test in GraphPad Prism software. P values of < 0.05 were considered statistically significant.
Results
We were able to demonstrate B cell lymphopenia in mice bone marrow downstream of pre-pro B cells, irrespective of pregnancy outcome. The mature bone marrow B cells did not show this adjustment mechanism during normal gestation.
Closer inspection of the splenic tissue revealed expansion and activation of marginal zone B cells in mice with a normal pregnancy. However, this was not observed in mice suffering from pregnancy disturbances. Natural antibodies secreted from marginal zone B cells were also present at higher concentrations in serum of pregnant mice, compared to non-pregnant animals.
We also found significantly higher levels of natural antibodies in serum of pregnant women compared to non-pregnant age-matched controls. Analysis showed significantly lower levels of BAFF in mice with normal pregnancy as compared to non-pregnant mice.
Conclusions
We are able to show mechanisms within the B cell compartment as well as the change within the natural antibodies that might be crucial for successful pregnancy in both humans and mice. Furthermore, BAFF seems to play a central role as a mediator of peripheral B cell compartment and B cell lymphopoiesis in the bone marrow for successful pregnancy.
Endothelial dysfunction (ED) comes with age, even without overt vessel damage such as that which occurs in atherosclerosis and diabetic vasculopathy. We hypothesized that aging would affect the downstream signalling of the endothelial nitric oxide (NO) system in the vascular smooth muscle (VSM). With this in mind, resistance mesenteric arteries were isolated from 13-week (juvenile) and 40-week-old (aged) mice and tested under isometric conditions using wire myography. Acetylcholine (ACh)-induced relaxation was reduced in aged as compared to juvenile vessels. Pretreatment with L-NAME, which inhibits nitrix oxide synthases (NOS), decreased ACh-mediated vasorelaxation, whereby differences in vasorelaxation between groups disappeared. Endothelium-independent vasorelaxation by the NO donor sodium nitroprusside (SNP) was similar in both groups; however, SNP bolus application (10−6 mol L−1) as well as soluble guanylyl cyclase (sGC) activation by runcaciguat (10−6 mol L−1) caused faster responses in juvenile vessels. This was accompanied by higher cGMP concentrations and a stronger response to the PDE5 inhibitor sildenafil in juvenile vessels. Mesenteric arteries and aortas did not reveal apparent histological differences between groups (van Gieson staining). The mRNA expression of the α1 and α2 subunits of sGC was lower in aged animals, as was PDE5 mRNA expression. In conclusion, vasorelaxation is compromised at an early age in mice even in the absence of histopathological alterations. Vascular smooth muscle sGC is a key element in aged vessel dysfunction.
Die vorliegende Studie basiert auf der Untersuchung von 89 verschiedenen Säugetierschädeln und 14 menschlichen Schädeln aus dem Institut für Anatomie und Zellbiologie der Universität Greifswald.
Die Arbeit vergleicht die Morphologie der knöchernen Schädel der Säugetiere mit denen des Menschen. Dabei wird von kieferorthopädischen Messpunkten, wie sie beim Menschen bereits seit längerer Zeit angewendet werden, ausgegangen. Sämtliche Messpunkte, die in der Kieferorthopädie herangezogen werden, finden sich auch an den Säugetierschädeln, sodass von einem gemeinsamen Schädelbauplan ausgegangen werden kann. Systematische Untersuchungen an einer größeren Gruppe von Säugetieren mit kieferorthopädischem Ansatz wurden, soweit ersichtlich, bisher noch nicht durchgeführt. Ziel dieser Arbeit ist es damit auch, eine dementsprechende Datengrundlage zu schaffen. Die dreidimensionale Vermessung erfolgte mit dem MicroScribe 3DX Digitalisierer (Immersion Corp., San Jose, CA) sowie einem digitalen Messschieber. Die Schädel wurden entsprechend der in der Literatur angewandten Taxonomie in Gruppen eingeteilt. Aus der Ordnung der Primaten wurden die Hominoidea, Platyrrhini, Cercopithecoidea und Lemuriformes untersucht, bei den Carnivora die Feliformia, Canidae, Ursidae und Pinnipedia. Bei den Cetartiodactyla wurden Ruminantia und Suidae vermessen, bei den Mesaxonia die Equidae.
In einem ersten Teil der Arbeit kommt die klassische Morphometrie zu Anwendung. Dabei werden klassische KFO- Messpunkte wie Nasion, Menton, Gonion, Pogonion, Zygion, Spina nasialis antetior, etc. verwendet, ebenso klassische Indizes wie Bolton-Analyse, Pont- Index und Izard- Index. In einem zweiten Teil kommt die geometrische Morphometrie zur Anwendung. Diese in der Biologie und Anthropologie bereits häufiger angewandte Methode wird jetzt auch vermehrt in der Kieferorthopädie angewandt. Durch die sogenannte Procrustes Transformation können dabei die vermessenen Schädel in Form und Gestalt unabhängig von der Größe verglichen werden. Bei sämtlichen Messungen werden die Unterschiede bzw. Gemeinsamkeiten zwischen Mensch und den einzelnen Säugetierarten herausgearbeitet und tabellarisch und graphisch dargestellt.
So können die beim Menschen nachgewiesenen kieferorthopädischen Indizes auch teilweise bei den Säugetieren gefunden werden. Sowohl beim Izard- Index als auch bei der Bolton- Analyse und der Tonn- Relation können Gemeinsamkeiten festgestellt werden. Größere Abweichungen gibt es dagegen beim Pont- Index, dem Gaumenhöhen- Index und dem Gaumen- Index. Auch der Jugomandibularindex zeigt wenig Übereinstimmung mit dem Menschen.
Dass die Primaten und hier insbesondere die Hominoidea und Cercopithecoidea dem Menschen schädelbezüglich am ähnlichsten sind, war zu erwarten und kann durch die hier vorliegenden Ergebnisse auch bestätigt werden. Allerdings weichen die Primaten bei der Gaumenform stärker vom Menschen ab, während bei der Gesichts- und Kieferform eine weniger zu erwartende Übereinstimmung mit Feliformia und Canidae festgestellt werden kann.
Das evolutionsbedingt stärkste Unterscheidungsmerkmal zu den Säugetieren ist der beim Menschen im Vergleich zur Schädellänge relativ kurze Gesichtsschädel.
Die in der Literatur beschriebene Taxonomie der Säugetiere, die durch eine Vielzahl verschiedener Untersuchungen hervorgegangen ist, kann hier sowohl mit Hilfe der klassischen Morphometrie als auch insbesondere durch die geometrische Morphometrie bestätigt werden. Bei der letzteren wird nach der Procrustes Transformation durch Clusterbildung sowohl nach deutlich voneinander abweichenden, als auch nach in sich homogenen Gruppen differenziert.
Durch die in dieser Studie differenziert herausgearbeiteten anatomische Strukturen in verschiedenen Schädelbereichen würde sich zusätzlich die Möglichkeit ergeben, bei kieferorthopädischen Tierversuchen in kraniofazialen Schädelregionen das anatomisch geeignete Tiermodell zu bestimmen, d.h. nicht eine einzelne Tierart ist für alle Versuche geeignet, sondern je nach Fragestellung müssen in unterschiedlichen Schädelbereichen verschiedene Tierarten herangezogen werden.
ABSTRACT
The Upper Pleistocene geoarchives in the south‐eastern Carpathian Basin are represented predominantly by loess–palaeosol records. In 2015, a 10 m sediment core composed of clay‐rich lacustrine sediments was recovered by vibracoring a dry lake basin located between the Vršac Mountains (Serbia) and the Banat Sands in the south‐eastern Carpathian Basin; a location relevant for placing regional archaeological results in a palaeoenvironmental context. Here, we present results from geoelectrical prospection and a lithostratigraphic interpretation of this sequence supported by a detailed granulometric study supplemented by ostracod analysis. An age model based on luminescence dating is discussed against sedimentological proxy data and its implication for palaeoenvironmental change. The cores show a stratigraphy of lighter ochre‐coloured and darker greyish sediment, related to the deposition of clay and silt trapped in an aquatic environment. Geophysical measurements show ~20 m thick lacustrine sediments. The grain‐size distributions including the variability in fine clay are indicative of a lacustrine environment. Fine particles were brought into the depositional environments by aquatic input and settled from suspension; also, direct dust input is constrained by grain‐size results. Riverine input and aeolian dust input interplayed at the locality.
Technological advances in light microscopy have always gone hand in hand with unprecedented biological insight. For microbiology, light microscopy even played a founding role in the conception of the entire discipline. The ability to observe pathogens that would otherwise evade human observation makes it a critical necessity and an indispensable tool to infectious disease research. Thus, the aim of this thesis was to optimize, extend, and functionally apply advanced light microscopy techniques to elucidate spatio-temporal and spatio-morphological components of bacterial and viral infection in vitro and in vivo.
Pathogens are in a constant arms race with the host’s immune system. By finding ways to circumvent host-mediated immune responses, they try to evade elimination and facilitate their own propagation. The first study (publication I) demonstrated that the obligate intracellular pathogen Coxiella burnetii is not just able to infect natural killer (NK) cells, but is actually capable of surviving the harsh degradative conditions in the cytotoxic lymphocyte’s granules. Using live-cell imaging of reporter-expressing Coxiella burnetii, the transient NK cell passage was closely monitored to provide detailed spatio-temporal information on this dynamic process in support of a range of static analyses. Bacterial release from NK cells was pinpointed to a time frame between 24 to 48 hours post-infection and the duration of release to about 15 minutes.
The second approach (publications II-V) aimed at shedding light on the greater spatio-morphological context of virus infection. Thus far, most studies investigating the distribution or tropism of viruses in vivo have used conventional immunohistochemistry in thin sections. Omitting the native spatial context of the infection site in vivo inherently bears the risk of incomplete description. While the microscopic tools and sample preparation protocols needed for volumetric 3D immunofluorescence imaging have recently been made available, they had not gained a foothold in virus research yet. An integral part of this thesis was concerned with the assessment and optimization of available tissue optical clearing protocols to develop an immunofluorescence-compatible 3D imaging pipeline for the investigation of virus infection inside its intact spatio-morphological environment (publication II). This formed the basis for all subsequent volumetric analyses of virus infection in vivo presented here. Consequently, this thesis provided a valuable proof of concept and blueprints for future virus research on the mesoscopic scale of host-pathogen interactions in vivo (publications II-V), using rabies virus (RABV; publications II-IV) and the newly-emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; publication V) as infection models for the nervous system and the respiratory tract, respectively.
Applying and further improving this volumetric 3D imaging workflow enabled unprecedented insights into the comprehensive in vivo cell tropism of RABV in the central (CNS) (publication III) and peripheral nervous system (PNS) (publication IV). Accordingly, differential infection of CNS-resident astrocytes by pathogenic and lab-attenuated RABV was demonstrated (publication III). While either virus variant showed equal capacity to infect neurons, as demonstrated by quantitative image analysis, only pathogenic field RABVs were able to establish non-abortive infection of astrocytes via the natural intramuscular inoculation route. A combined 3D LSFM-CLSM workflow further identified peripheral Schwann cells as a relevant target cell population of pathogenic RABV in the PNS (publication IV). This suggested that non-abortive infection of central and peripheral neuroglia by pathogenic RABV impairs their immunomodulatory function and thus represents a key step in RABV pathogenesis, which may contribute significantly to the establishment of lethal rabies disease.
Finally, utilizing the full volumetric acquisition power of LSFM, a further refined version of the established 3D imaging pipeline facilitated a detailed mesoscopic investigation of the distribution of SARS-CoV-2 in the respiratory tract of the ferret animal model (publication V). Particularly for this newly-emerged pathogen of global concern, in-depth knowledge of host-pathogen interactions is critical. By preserving the complete spatio-morphological context of virus infection in the ferret respiratory tract, this thesis provided the first specific 3D reconstruction of SARS-CoV-2 infection and the first report of 3D visualization of respiratory virus infection in nasal turbinates altogether. 3D object segmentation of SARS-CoV-2 infection in large tissue volumes identified and emphasized a distinct oligofocal infection pattern in the upper respiratory tract (URT) of ferrets. Furthermore, it corroborated a preferential replication of SARS-CoV-2 in the ferret URT, as only debris-associated virus antigen was detected in the lower respiratory tract of ferrets, thus providing crucial information on the spatial distribution of SARS-CoV-2.
Objective: To compare the effectiveness and complications of intraligamentary anesthesia
(ILA) with conventional inferior alveolar nerve block (IANB) during injection and dental
treatment of mandibular posterior teeth.
Materials and Methods: In this randomized, prospective clinical trial, 72 patients (39 males, 33
females) patients scheduled for dental treatment of mandibular posterior teeth, were randomly
allocated to ILA group (n=35) received ILA injection or IANB group (n=37) received the
conventional IANB. Our primary outcome was to assess pain and stress (discomfort) during the
injection and dental treatment, using the Numeric Rating Scale (NRS) from 0 to 10 (0 = no
pain, 10= the worst pain imaginable). Whereas; recording 24 hours postoperative complications
were our Secondary outcomes.
Results: Patients in ILA group reported significantly less pain during injection when compared
with IANB group (p=0.03). While pain during dental treatment was similar in both groups
(p=0.2). Patients in both groups also reported similar low values of discomfort during treatment
(p= 0.7). Although no signs of nerve contact or any other postoperative complications were
observed, five patients in IANB group (none in ILA group) reported temporary irritations
Conclusion: This study showed equivalent effectiveness of both intraligamentary anesthesia
and conventional inferior alveolar nerve block, for pain control during routine dental treatment
of mandibular posterior teeth. Nevertheless, ILA showed significantly less pain during
injection. No major postoperative complications in both groups were observed.
Clinical Relevance: ILA could be considered as an effective alternative for routine dental
treatment.
Polyethylene terephthalate (PET) is a mass-produced petroleum-based non-biodegradable plastic that contributes to the global plastic pollution. Recently, biocatalytic degradation has emerged as a viable recycling approach for PET waste, especially with thermophilic polyester hydrolases such as a cutinase (LCC) isolated from a leaf-branch compost metagenome and its variants. To improve the enzymatic PET hydrolysis performance, we fused a chitin-binding domain (ChBD) from Chitinolyticbacter meiyuanensis SYBC-H1 to the C-terminus of the previously reported LCCICCG variant, demonstrating higher adsorption to PET substrates and, as a result, improved degradation performance by up to 19.6% compared to with its precursor enzyme without the binding module. For compare hydrolysis with different binding module, the catalytic activity of LCCICCG-ChBD, LCCICCG-CBM, LCCICCG-PBM and LCCICCG-HFB4 were further investigated with PET substrates of various crystallinity and it showed measurable activity on high crystalline PET with 40% crystallinity. These results indicated that fusing a polymer-binding module to LCCICCG is a promising method stimulating the enzymatic hydrolysis of PET.
Sturgeons are among the most ancient linages of actinopterygians. At present, many sturgeon species are critically endangered. Surrogate production could be used as an affordable and a time-efficient method for endangered sturgeons. Our study established a method for identifying and isolating type A spermatogonia from different developmental stages of testes using flow cytometric cell sorting (FCM). Flow cytometric analysis of a whole testicular cell suspension showed several well-distinguished cell populations formed according to different values of light scatter parameters. FCM of these different cell populations was performed directly on glass slides for further immunocytochemistry to identify germ cells. Results showed that the cell population in gate P1 on a flow cytometry plot (with high forward scatter and high side scatter parameter values) contains the highest amount of type A spermatogonia. The sorted cell populations were characterized by expression profiles of 10 germ cell specific genes. The result confirmed that setting up for the P1 gate could precisely sort type A spermatogonia in all tested testicular developmental stages. The P2 gate, which was with lower forward scatter and side scatter values mostly, contained type B spermatogonia at a later maturing stage. Moreover, expressions of plzf, dnd, boule, and kitr were significantly higher in type A spermatogonia than in later developed germ cells. In addition, plzf was firstly found as a reliable marker to identify type A spermatogonia, which filled the gap of identification of spermatogonial stem cells in sterlet. It is expected to increase the efficiency of germ stem cell culture and transplantation with plzf identification. Our study thus first addressed a phenotypic characterization of a pure type A spermatogonia population in sterlet. FCM strategy can improve the production of sturgeons with surrogate broodstock and further the analysis of the cellular and molecular mechanisms of sturgeon germ cell development.
Background: Gastrointestinal hormones (GIHs) are crucial for the regulation of a variety of physiological functions and have been linked to hunger, satiety, and appetite control. Thus, they might constitute meaningful biomarkers in longitudinal and interventional studies on eating behavior and body weight control. However, little is known about the physiological levels of GIHs, their intra-individual stability over time, and their interaction with other metabolic and lifestyle-related parameters. Therefore, the aim of this pilot study is to investigate the intra-individual stability of GIHs in normal-weight adults over time. Methods: Plasma concentrations of ghrelin, leptin, GLP-1 (glucagon-like-peptide), and PP (pancreatic polypeptide) were assessed by enzyme-linked immunosorbent assay (ELISA) in 17 normal-weight, healthy adults in a longitudinal design at baseline and at follow-up six months later. The reliability of the measurements was estimated using intra-class correlation (ICC). In a second step, we considered the stability of GIH levels after controlling for changes in blood glucose and hemoglobin A1 (HbA1c) as well as self-reported physical activity and dietary habits. Results: We found excellent reliability for ghrelin, good reliability for GLP1 and PP, and moderate reliability for leptin. After considering glucose, HbA1c, physical activity, and dietary habits as co-variates, the reliability of ghrelin, GLP1, and PP did not change significantly; the reliability of leptin changed to poor reliability. Conclusions: The GIHs ghrelin, GLP1, and PP demonstrated good to excellent test–retest reliability in healthy individuals, a finding that was not modified after adjusting for glucose control, physical activity, or dietary habits. Leptin showed only moderate to poor reliability, which might be linked to weight fluctuations, albeit small, between baseline and follow-up assessment in our study sample. Together, these findings support that ghrelin, GLP1, and PP might be further examined as biomarkers in studies on weight control, with GLP1 and PP serving as anorexic markers and ghrelin as an orexigenic marker. Additional reliability studies in obese individuals are necessary to verify or refute our findings for this cohort.
Pentathiepins are polysulfur-containing compounds that exert antiproliferative and cytotoxic activity in cancer cells, induce oxidative stress and apoptosis, and inhibit glutathione peroxidase (GPx1). This renders them promising candidates for anticancer drug development. However, the biological effects and how they intertwine have not yet been systematically assessed in diverse cancer cell lines. In this study, six novel pentathiepins were synthesized to suit particular requirements such as fluorescent properties or improved water solubility. Structural elucidation by X-ray crystallography was successful for three derivatives. All six underwent extensive biological evaluation in 14 human cancer cell lines. These studies included investigating the inhibition of GPx1 and cell proliferation, cytotoxicity, and the induction of ROS and DNA strand breaks. Furthermore, selected hallmarks of apoptosis and the impact on cell cycle progression were studied. All six pentathiepins exerted high cytotoxic and antiproliferative activity, while five also strongly inhibited GPx1. There is a clear connection between the potential to provoke oxidative stress and damage to DNA in the form of single- and double-strand breaks. Additionally, these studies support apoptosis but not ferroptosis as the mechanism of cell death in some of the cell lines. As the various pentathiepins give rise to different biological responses, modulation of the biological effects depends on the distinct chemical structures fused to the sulfur ring. This may allow for an optimization of the anticancer activity of pentathiepins in the future.
The genus Capripoxvirus of the family Poxviridae consists of the species lumpy skin disease virus, sheeppox virus and goatpox virus that affect cattle, sheep and goats, respectively. Whereas lumpy skin disease virus (LSDV) is transmitted mainly mechanically via blood-feeding insects and possibly hard ticks, the major transmission routes of sheeppox virus (SPPV) and goatpox virus (GTPV) are via direct contact and aerosols. Affected animals develop fever and display clinical signs such as ocular and nasal discharge, lymphadenopathy and characteristic lesions of the skin. Severe clinical course, especially in combination with respiratory signs, can result in the death of the affected animals. In endemic regions, mortality of capripox virus-induced diseases is low (1-10%). However, mortalities of up to 75% have been reported for LSDV and up to 100% for SPPV and GTPV in exotic breeds and high-producing dairy or beef animals. The loss of quality of the leather, reduced weight gain and milk yield as well as complete loss of affected animals have severe impact on national and global economies. Therefore, capripox virus-induced diseases have significant impact on both the affected individual animal as well as on the existence of small-scale farmers and large agricultural enterprises. However, until now, only live attenuated vaccines are commercially available. These attenuated vaccines are not authorized in the European Union and their administration would comprise the disease-free status of the respective country. Thus, reliable diagnostic tools for the detection and characterization of capripox viruses as well as safe and efficient control measures are of high importance.
The objectives of the present thesis were the development, validation and comparison of diagnostic tools, the establishment of challenge infection models and the performance of pathogenesis studies for all three capripox virus species, and the development and testing of different inactivated prototype vaccine candidates against LSDV.
First, new real-time quantitative polymerase chain reaction (qPCR) assays for robust detection and differentiation of LSDV field strains, LSDV vaccine strains, SPPV and GTPV were developed and extensively validated. In the following, two single assays were combined to duplex assays, one for the differentiation between LSDV field strains and LSDV vaccine strains, and the second for discrimination of SPPV and GTPV. Finally, a diagnostic workflow based on these new duplex assays in combination with already published methods was established. This workflow enables time-saving, robust and reliable detection, species-specific identification and genetic and phylogenetic characterization of all three capripox virus species. In addition, already existing serological examination methods (serum neutralization assay and commercial enzyme-linked immunosorbent assay) were compared regarding their sensitivity and specificity. Furthermore, pathogenesis studies with different capripox virus isolates were performed in the respective target species, and the suitability of selected virus isolates as challenge viruses for future vaccine studies was analyzed. Pathogenesis studies with isolates GTPV-“V/103” and LSDV-“Macedonia2016” revealed that both are proper candidates for challenge models. Finally, three different SPPV isolates (SPPV-“V/104”, SPPV-“India/2013/Surankote” and SPPV-“Egypt/2018”) were tested in sheep regarding their virulence to find a suitable challenge model for SPPV, and SPPV-“India/2013/Surankote” was chosen for future vaccine studies.
Once appropriate challenge models were established, different inactivated prototype vaccines against LSDV were developed, and vaccine safety as well as vaccine efficacy were tested in cattle. Eventually, a Polygen-adjuvanted inactivated LSDV-vaccine candidate was selected that is able to fully prevent cattle from any LSDV-related clinical signs after severe challenge infection. Furthermore, molecular and serological data indicate that this inactivated prototype vaccine is even able to induce a kind of “sterile immunity” against LSDV in those cattle. It has to be mentioned that a commercially available vaccine similar to this prototype vaccine would be a great advance for the control of LSDV.
In the future, additional studies addressing diagnostics and optimized control of capripox viruses should be performed. Firstly, probe-based real-time qPCR assays for the differentiation of SPPV and GTPV vaccine strains from their respective virulent field strains should be developed and included into the diagnostic workflow. Secondly, further tests of the inactivated prototype vaccine, e.g. determination of the minimum protective dose and the possibility of cross-protection in sheep and goats against SPPV and GTPV, respectively, should be performed.
Es wurden jeweils 50 Patienten mit antagonistenlosen Molaren und Prämolaren sowie mit Vollbezahnung befragt und untersucht. Dabei wurde eine umfangreiche Befund- und Funktionserhebung durchgeführt, die Elongation der betroffenen Zähne ermittelt, sowie gesellschaftlich-soziale Parameter und subjektive Symptome wie Schmerzen, Kaugeräusche und sonstige Missempfindungen abgefragt. Mittels OHIP wurde die MLQ ermittelt.
In der Gruppe der Patienten mit antagonistenlosen Molaren und Prämolaren konnte bei 18% der Zähne keine Elongation, bei 22% eine leichte Elongation von weniger als 2 mm und bei 59% eine stark ausgeprägte Elongation von 2 mm und mehr festgestellt werden.
Ferner war in der Gruppe der Patienten mit antagonistenlosen Molaren und Prämolaren die Chance erhöht, an Schmerzen und Funktionsstörungen zu leiden sowie eine schlechtere MLQ zu haben. Die Ergebnisse zeigen eine Tendenz, die als moderate Assoziation nachgewiesen werden kann. Für die Unterschiede gelingt der Nachweis der statistischen Signifikanz nicht.
Für weitere Studien sind multizentrische Ansätze mit deutlich höheren Stichprobenumfängen (>300) als in dieser Arbeit und einem multidimensionalen Untersuchungsdesign zu empfehlen, deren umfangreiche strukturelle Anforderungen eine besondere Herausforderung sein werden. Letztlich zeigt auch diese Arbeit, dass jede antagonistenlose Situation eine individuelle Entscheidung anhand der klinischen Gegebenheiten verlangt.
Eine Vielzahl von Faktoren kann die Entwicklung des Kindes vor und nach der Geburt
beeinflussen. Um diese Faktoren zu detektieren, wurde die Studie Survey of
Neonates in Pomerania, im Nordosten von Deutschland, initiiert.
Von 2002 bis 2008 fand die Basisuntersuchung (SNiP-I) statt. Auf die gute Teilnehmerrate
von 75% konnte mit der Nachuntersuchung (SNiP-II-Follow-up) aufgebaut
und die Querschnittsstudie zu einer Längsschnittstudie umgewandelt werden. Die
nun 9- bis 16-jährigen Kinder und Jugendlichen und deren Eltern wurden erneut zu
Themen der körperlichen und seelischen Gesundheit, zu sozioökonomischen- und
Umweltfaktoren in Form eines selbstauszufüllenden Fragebogens befragt. Es wurden
im Gegensatz zur SNiP-I Untersuchung keine Bioproben genommen.
Die Nachuntersuchung fand von Dezember 2016 bis Juli 2017 statt. Es wurde eine
Wiederteilnahmerate von 28.8% (1665 von 5725) und eine Deckungsrate von 20%
erreicht. Wie in anderen Nachuntersuchungen im Rahmen von Geburtskohorten ist
eine Verschiebung zu höherem sozioökonomischen Status festgestellt worden: die
Frauen waren älter, gebildeter und hatten ein höheres Einkommen.
Die akute Pankreatitis ist eine der häufigsten nicht malignen gastrointestinalen Erkrankungen, die zu Krankenhausaufenthalten führt. Sie ist als Selbstverdau des Pankreas durch seine eigenen Proteasen wie z.B. Trypsin, Elastase und Chymotrypsin definiert. Als Ursprung der Erkrankung wird die frühzeitige intrazelluläre Aktivierung dieser Verdauungsenzyme angesehen. Dies führt zum Zelltod der Azinuszellen und zur Schädigung des Gewebes.
Während der akuten Pankreatitis kommt es in 20% der Fälle zu einem schweren Verlauf der Erkrankung, der mit Organversagen in der Lunge und den Nieren assoziiert ist. Es ist bekannt, dass es zu einer Entzündungsreaktion kommt, bei der große Mengen an Zytokinen ausgeschüttet werden. Leukozyten infiltrieren das Pankreas und verstärken den Gewebeschaden. Es kommt zur Freisetzung von DAMPs, die das angeborene und adaptive Immunsystem aktivieren. Bislang ist nicht gut untersucht, wie das Immunsystem den schweren Verlauf der akuten Pankreatitis beeinflusst und es gibt wenig Theorien über den Organschaden in der Lunge und den Nieren.
In dieser Arbeit lag der Fokus auf dem Organschaden in Lunge und Niere und die Wirkung von Interleukin 33 (IL33) auf die Zellen des angeborenen Immunsystems und deren Einwanderung in verschiedene Organe während der schweren akuten Pankreatitis im Mausmodell. Die schwere akute Pankreatitis wurde mittels Gangligatur und einmaliger Gabe von Caerulein an Tag 2 nach Gangligatur induziert. An Tag 3 nach Induktion wurden die Mäuse getötet und die Organe wurden für weitere Analysen entnommen.
Am dritten Tag nach Induktion der Pankreatitis kam es zu einem Organschaden in der Lunge und den Nieren. In der Lunge fand sich eine Verdickung der Alveolarsepten und eine Verdichtung des Gewebes sowie eine Infiltration von Leukozyten und ein Ödem. In der Niere waren ebenfalls strukturelle Veränderungen zu finden und eine Infiltration von Leukozyten war zu beobachten. In durchflusszytometrischen Analysen der Lunge konnte beobachtet werden, dass CD11b+CD62L+ Monozyten während der akuten Pankreatitis signifikant anstiegen. Mittels RT-DC wurde gezeigt, dass diese Monozyten an Tag 3 signifikant an Größe zugenommen hatten. Mit einer CD11b Färbungen von Lungen und Nieren konnte die Infiltration durch Monozyten bestätigt werden. Unter einer Blockade von Monozyten durch systemische Gabe von anti-CCR2-Antikörpern verringerte sich die Schädigung in Lunge und Niere während der Pankreatitis signifikant.
Diese Daten legen nahe, dass der Organschaden in der schweren akuten Pankreatitis durch infiltrierende Monozyten verursacht wird, die über CD62L (L-Selektin) an die Gefäßwände binden und über ihre Größe Gefäße verstopfen, was in den Kapillaren zur Ischämie führt.
In vitro sezernierten Makrophagen, die mit CCK stimulierten Azinuszellen co-inkubiert wurden, IL33. Im Mausmodell wurde IL33 mittels sST2 blockiert, was die Schädigung des Pankreas in der Pankreatitis reduzierte. In IL33-depletierten Tieren fand sich im Vergleich zum Wildtyp ein geringerer Lungenschaden aber eine unveränderte Nierenschädigung. Somit scheint IL33 eine Rolle bei der Monozyten-vermittelten Organschädigung in der Pankreatitis zu spielen, die sich auf Grund von kompensatorischen Regulationsmechanismen im globalen IL33 Knock-out weniger gut belegen lässt als nach IL33 Inhibition. Die Hemmung von IL33 zur Behandlung der akuten Pankreatitis stellt somit ein vielversprechendes Therapieprinzip dar.