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Background: Gram-negative infections of the peritoneal cavity result in profound modifications of peritoneal B cell populations and induce the migration of peritoneal B cells to distant
secondary lymphoid organs. However, mechanisms controlling the egress of peritoneal B cells from
the peritoneal cavity and their subsequent trafficking remain incompletely understood. Sphingosine1-phosphate (S1P)-mediated signaling controls migratory processes in numerous immune cells. The
present work investigates the role of S1P-mediated signaling in peritoneal B cell trafficking under
inflammatory conditions. Methods: Differential S1P receptor expression after peritoneal B cell activation was assessed semi-quantitatively using RT-PCR in vitro. The functional implications of
differential S1P1 and S1P4 expression were assessed by transwell migration in vitro, by adoptive
peritoneal B cell transfer in a model of sterile lipopolysaccharide (LPS)-induced peritonitis and in
the polymicrobial colon ascendens stent peritonitis (CASP) model. Results: The two sphingosine-1-
phosphate receptors (S1PRs) expressed in peritoneal B cell subsets S1P1 and S1P4 are differentially
regulated upon stimulation with the TLR4 agonist LPS, but not upon PMA/ionomycin or B cell receptor (BCR) crosslinking. S1P4 deficiency affects both the trafficking of activated peritoneal B cells
to secondary lymphoid organs and the positioning of these cells within the functional compartments of the targeted organ. S1P4 deficiency in LPS-activated peritoneal B cells results in significantly reduced numbers of splenic innate response activator B cells. Conclusions: The S1P-S1PR system is implicated in the trafficking of LPS-activated peritoneal B cells. Given the protective role of peritoneal B1a B cells in peritoneal sepsis, further experiments to investigate the impact of S1P4 mediated signaling on the severity and mortality of peritoneal sepsis are warranted.
Abstract
(1) Background: Surgery is the most important element of multimodal treatment concepts in oncological patients, especially in the early stages of pancreatic tumours. While the influence of primary tumour resection on the immune status was analysed in several studies, the impact of tumour-unrelated visceral surgery on the tumour-bearing organism and on the primary tumour itself is not yet fully understood. (2) Methods: We combined a murine model of orthotopically implanted adenocarcinoma of the pancreas with the model of surgically-induced immune dysfunction (SID). Mortality and general condition including body weight were observed over a period of 28 days. Tumour growth was analysed by MRI scans on days 8 and 27 following tumour implantation. On day 28, the immune cell populations in the blood and spleen as well as the serum cytokines were quantified. (3) Results: SID results in a significant deterioration of the general condition and a reduced increase in the body weight of tumour-bearing mice compared to the control groups, while mortality and tumour growth rate were not influenced. The numbers of spleen macrophages and neutrophils were increased in tumour-bearing animals following SID. Furthermore, both macrophage and neutrophil levels were increased in the peripheral blood. (4) Conclusions: The presented results might contribute to the basic understanding of the interaction of tumour and immune system and could contribute to new approaches to immunotherapeutic strategies.
Structured examination and treatment are essential in medicine. For dental students, a structured approach to the assessment of oral mucosal lesions is missing thus far. To validate an approach, a structured questionnaire was compared with the habitually used free description of oral lesions (white lesions, ulcers, hyperplasia). Thirty-three dental students were divided into two groups (Group 1 (n = 17) used the free description; Group 2 (n = 16) used a guided questionnaire) to characterize mucosal lesions in patients and make a tentative diagnosis. Although no difference was found between the groups regarding the suspected diagnosis or the histopathological findings, there was a significant advantage of the structured questionnaire in all aspects of the description compared to the free description (p = 0.000018). Thus, a structured description is an important aspect in the evaluation of oral mucosal changes, and a guided questionnaire should be implemented in the study of dentistry.
Reactive species generated by medical gas plasma technology can be enriched in liquids for use in oncology targeting disseminated malignancies, such as metastatic colorectal cancer. Notwithstanding, reactive species quantities depend on the treatment mode, and we recently showed gas plasma exposure in conductive modes to be superior for cancer tissue treatment. However, evidence is lacking that such a conductive mode also equips gas plasma-treated liquids to confer augmented intraperitoneal anticancer activity. To this end, employing atmospheric pressure argon plasma jet kINPen-treated Ringer’s lactate (oxRilac) in a CT26-model of colorectal peritoneal carcinomatosis, we tested repeated intraabdominal injection of such remotely or conductively oxidized liquid for antitumor control and immunomodulation. Enhanced reactive species formation in conductive mode correlated with reduced tumor burden in vivo, emphasizing the advantage of conduction over the free mode for plasma-conditioned liquids. Interestingly, the infiltration of lymphocytes into the tumors was equally enhanced by both treatments. However, significantly lower levels of interleukin (IL)4 and IL13 and increased levels of IL2 argue for a shift in intratumoral T-helper cell subpopulations correlating with disease control. In conclusion, our data argue for using conductively over remotely prepared plasma-treated liquids for anticancer treatment.
Gas plasma is an approved technology that generates a plethora of reactive oxygen species, which are actively applied for chronic wound healing. Its particular antimicrobial action has spurred interest in other medical fields, such as periodontitis in dentistry. Recent work has indicated the possibility of performing gas plasma-mediated biofilm removal on teeth. Teeth frequently contain restoration materials for filling cavities, e.g., resin-based composites. However, it is unknown if such materials are altered upon gas plasma exposure. To this end, we generated a new in-house workflow for three commonly used resin-based composites following gas plasma treatment and incubated the material with human HaCaT keratinocytes in vitro. Cytotoxicity was investigated by metabolic activity analysis, flow cytometry, and quantitative high-content fluorescence imaging. The inflammatory consequences were assessed using quantitative analysis of 13 different chemokines and cytokines in the culture supernatants. Hydrogen peroxide served as the control condition. A modest but significant cytotoxic effect was observed in the metabolic activity and viability after plasma treatment for all three composites. This was only partially treatment time-dependent and the composites alone affected the cells to some extent, as evident by differential secretion profiles of VEGF, for example. Gas plasma composite modification markedly elevated the secretion of IL6, IL8, IL18, and CCL2, with the latter showing the highest correlation with treatment time (Pearson’s r > 0.95). Cell culture media incubated with gas plasma-treated composite chips and added to cells thereafter could not replicate the effects, pointing to the potential that surface modifications elicited the findings. In conclusion, our data suggest that gas plasma treatment modifies composite material surfaces to a certain extent, leading to measurable but overall modest biological effects.
(1) Background: The aim of this study was to systematically compare TEM sections of mineralized human enamel and dentine prepared by focused ion beam (in situ lift-out) technique and ultramicrotomy through a combination of microscopic examination methods (scanning electron microscopy and transmission electron microscopy). In contrast with published studies, we compared the TEM preparation methods using the same specimen blocks as those for the ultramicrotomy and FIB technique. (2) Methods: A further evaluation of TEM sample preparation was obtained by confocal laser scanning microscopy and atomic force microscopy. In addition, ultramicrotome- and focused ion beam-induced artefacts are illustrated. (3) Results: The FIB technique exposed a major difference between non-decalcified enamel and dentine concerning the ultrastructural morphology compared to ultramicrotome-prepared sections. We found that ultramicrotomy was useful for cutting mineralized dentine, with the possibility of mechanical artefacts, but offers limited options for the preparation of mineralized enamel. FIB preparation produced high-quality TEM sections, showing the anisotropic ultrastructural morphology in detail, with minor structural artefacts. Our results show that the solution of artificial saliva and glutardialdehyde (2.5% by volume) is a very suitable fixative for human mineralized tissue. (4) Conclusions: The protocol that we developed has strong potential for the preparation of mineralized biomaterials for TEM imaging and analysis.
Plasma medical oncology: Immunological interpretation in head and neck squamous cell carcinoma
(2020)
Over the past several years, various important articles focusing on cancer therapy
approaches in head and neck squamous cell carcinomas (HNSCCs) using cold
atmospheric plasma (CAP) have been published (SEMMLER et al. 2020 [53],
METELMANN et al. 2018 [44], KEIDAR et al. 2011 [33]). This doctoral thesis presents
selected results from a prospective observational clinical study in CAP therapy of
palliative HNSCC patients, carried out at the Department of Oral and Maxillofacial
Surgery/Plastic Surgery of the Greifswald University Medicine. For oral and
maxillofacial surgeons, ulcerated surfaces of locally advanced head and neck squamous
cell carcinomas (UICC IV) offer a challenging treatment assignment with microbial
contamination and tumour progression. The clinical attempt appears to eradicate
microbial contamination and to initiate tumour regression. This doctoral thesis will
describe the processes of human tumour biology and tumour immunology in HNSCCs
and the extent of present knowledge concerning plasma medical oncology as an
anticancer modality. In the introduction of the doctoral thesis clinical results of plasma
therapy in locally advanced HNSCCs (UICC IV) are set out. This mainly includes the
investigation of a therapeutic concept, the treatment phases, the tumour size development
and the morphological changes of the infected tumour surface following cold atmospheric
plasma therapy. In the main part, a detailed immunological interpretation is proposed on
the basis of present preclinical and clinical immunological knowledge. Finally,
unexplored questions in plasma medical oncology are highlighted. This is highly
significant for future plasma research and clinical anticancer therapy.
In der vorliegenden Arbeit wurde eine Smartphone-App mit einem Papierfragebogen zur Selbstbeobachtung bei Craniomandibulären Dysfunktionen (CMD) verglichen. Das primäre Ziel dieser Studie war die Gegenüberstellung der Interventionen „CMD-App“ und „Papierfragebogen“ und die Untersuchung der Probandencompliance bei deren Anwendung. Sekundär sollten Alltagsintegrität, Übersichtlichkeit, Achtsamkeit und Zeitaufwand der Interventionen untersucht werden. Dazu nahmen in einem Zeitraum von 20 Monaten insgesamt 81 Probanden, im Alter von 21 bis 36 Jahren, an der Studie teil. Die Studienpopulation bestand aus Zahnmedizinstudenten der Universitätsmedizin Greifswald im klinischen Studienabschnitt. Die Selbstbeobachtung wurde von beiden Gruppen vier Tage lang jeweils fünf Mal täglich durchgeführt. Für diese Untersuchung wurden die verwendeten Interventionen von uns neu entwickelt. Die Papierfragebogengruppe nutzte im gesamten Beobachtungszeitraum vier Papierfragebögen in Gestaltung und Format eines Smartphones zusammen mit roten Klebepunkten als Erinnerungshilfe. Die Appgruppe benutzte eine Android-Smartphoneapp mit dem Namen „CMD-App“, welche als Erinnerung an die Selbstbeobachtung Push-Benachrichtigungen versendete. Die vorliegende Arbeit diente als erste Pilotuntersuchung für die „CMD-App“.
Die Ergebnisse dieser Studie zeigten, dass die Probandencompliance bei Nutzern der „CMD-App“ höher war als bei Probanden der Papierfragebogengruppe. Gemessen an der Gesamtanzahl an Eintragungen zur Selbstbeobachtung, war die „CMD-App“ dem Papierfragebogen überlegen. Die Probanden in der Appgruppe haben durchschnittlich 8,9 % mehr Selbstbeobachtungen durchgeführt als die Probanden, die den Papierfragebogen nutzten. In den Punkten Alltagsintegrität, Übersichtlichkeit und Zeitaufwand war die „CMD-App“ im Vergleich zum Papierfragebogen ebenfalls im Vorteil. Es konnte auch gezeigt werden, dass 76,67 % der Probanden der Appgruppe durchschnittlich nur 2 Minuten pro Tag mit der Selbstbeobachtung verbrachten.
In weiteren klinischen Studien gilt es nun zu klären, ob sich die von uns erhobenen Ergebnisse dieser Pilotuntersuchung auch auf CMD-Patienten übertragen lassen. Diese Arbeit konnte bereits einen Hinweis darauf geben, wie digitale Medien in der Zahnheilkunde in Zukunft genutzt werden könnten. Eine technisch ausgereifte „CMD-App“ hat Potenzial Zahnärzte in der Diagnose und Therapie von CMD zu unterstützen und die herkömmliche Methode zur Selbstbeobachtung abzulösen oder zu ergänzen.
The Study of Health in Pomerania (SHIP), a population-based study from a rural state in northeastern Germany with a relatively poor life expectancy, supplemented its comprehensive examination program in 2008 with whole-body MR imaging at 1.5 T (SHIP-MR). We reviewed more than 100 publications that used the SHIP-MR data and analyzed which sequences already produced fruitful scientific outputs and which manuscripts have been referenced frequently. Upon reviewing the publications about imaging sequences, those that used T1-weighted structured imaging of the brain and a gradient-echo sequence for R2* mapping obtained the highest scientific output; regarding specific body parts examined, most scientific publications focused on MR sequences involving the brain and the (upper) abdomen. We conclude that population-based MR imaging in cohort studies should define more precise goals when allocating imaging time. In addition, quality control measures might include recording the number and impact of published work, preferably on a bi-annual basis and starting 2 years after initiation of the study. Structured teaching courses may enhance the desired output in areas that appear underrepresented.
Die autologe Fettgewebstransplantation ist eine in der plastischen Chirurgie routinemäßig angewandte Methode. Die Indikationen reichen von der Rekonstruktion nach Teilresektion der Mamma, über Einspritzung bei narbigen Einziehungen zur Wiederherstellung des Körperreliefs, bis zum Einsatz bei Lipodystrophien wie sie beispielsweise im Rahmen von HIV-Medikationen vorkommen können. Dieser Eingriff erfolgt überwiegend in Lokalästhesie, unter Verwendung von Lidocain, um dem Patienten die Belastung einer Vollnarkose zu ersparen. Die Lipotoxizität dieses Lokalanästhetikums würde hierbei dessen Einsatz ausschließen. Die vorliegende Studie verwendet einen Versuchsaufbau, der es ermöglicht bei Patienten intraindividuell den Einfluss von Lidocain auf Zellen des Fettgewebes quantitativ zu evaluieren. Nach Zellisolierung aus dem Gewebeverband wurden einzelne Zelltypen der Stromal-Vascular Fraction mittels Antikörperfärbung identifiziert. Wir konnten zeigen, dass Lidocain keinen Einfluss auf prozentuale Verteilung, Absolutzellzahlen und Vitalität von Fettgewebsstammstellen, Präadipozyten, reifen Fettzellen und Leukozyten hat. Zudem unterstützen die Ergebnisse, dass beim Lipotransfer ausschließlich die feste Phase des Lipoaspirates transplantiert werden sollte. Insgesamt sprechen die Ergebnisse der vorliegenden Studie für die Verwendung von Lidocain in therapeutischer Konzentration zur Eigenfetttransplantation, was gegen die Notwendigkeit einer Vollnarkose spricht. Die gewählte Methodik verwendet einen Versuchsaufbau der Daten mit direkter Relevanz für die Durchführung der Eigenfetttransplantation generiert. Sie leistet somit einen wichtigen Beitrag zur Optimierung des Lipotransfers. Dies schmälert die Notwendigkeit von Folgeeingriffen zur Revision, was unter anderem zur Steigerung der Patientenzufriedenheit führt.
Bei der Vielzahl der bestehenden Methoden wäre für zukünftige Forschung in dem Bereich der Eigenfetttransplantation eine genauere Abstimmung der einzelnen Forschungsgruppen untereinander nötig. Für weitere Studien sind somit von wirtschaftlichen Interessen unabhängige und international koordinierte Untersuchungen wünschenswert.