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Purpose: The cyclin-dependent kinase (Cdk) inhibitor p27Kip1 may be involved in regulating re-entry of residual hepatocytes into the cell cycle upon loss of liver tissue by partial hepatectomy (PH). As yet, changes in Kip1 expression during the initial period following PH are not well-characterized. We investigated immediate changes in Kip1 mRNA and protein levels as well as changes in Kip1 phosphorylation in liver tissue within the relevant time window between surgery and the onset of DNA synthesis at 10–12 h.
Methods: We used real-time PCR, quantitative Western blotting, and immune histochemistry on tissue samples of adult rats obtained during or between 2 and 10 h after surgical removal of two thirds of the liver to analyze Kip1 mRNA or protein levels, respectively, or to quantify nuclear expression of Kip1.
Results: Kip1 mRNA was down-regulated within 4 h after PH by 60% and remained unchanged thereafter up to 10 h. With a lag phase of 2–3 h, Kip1-protein was down-regulated to a level of 40% of the control. The level of Thr187-phosphorylated Kip1 started to increase at 4 h and reached a maximum level at 8–10 h after PH. Kip1 immunoreactivity was observed in 30% of the hepatocytes before PH. Within 6–8 h after PH, more than half of the hepatocytes lost nuclear Kip1 signals. Kip1-specific micro-RNAs (miRNA221, miRNA222) were not changed upon PH.
Conclusions: A portion of hepatocytes in adult rats constitutively express Kip1 and down-regulate Kip1 immediately upon PH. This response involves transcriptional processes (loss of Kip1 mRNA) as well as accelerated degradation of existing protein (increase in pThr187-phosphorylation mediating polyubiquitinylation and proteasomal degradation of Kip1). Kip1 down-regulation occurs precisely within the intervall between surgery and onset of DNA synthesis which supports the hypothesis that it mediates activation of G0/0S-phase Cdk/cyclin-complexes and re-entry of hepatocytes into the cell cycle.
Background: Patients with mucin-producing adenocarcinoma have an increased risk for venous and arterial thrombosis. When these patients present with thrombocytopenia, disseminated intravascular coagulopathy (DIC) is often the underlying cause. Case Report: We report 2 patients who were admitted due to bleeding symptoms of unknown cause, in whom further workup revealed adenocarcinoma-induced DIC. Conclusion: In elderly patients presenting with signs of DIC, such as reduced fibrinogen levels, elevated prothrombin time, elevated D-dimer, and thrombocytopenia, without any obvious reason (e.g., sepsis), adenocarcinoma-associated coagulopathy should be considered as the underlying cause. Paradoxically, in these patients bleeding symptoms improve when the patient is sufficiently anti-coagulated with low molecular weight heparin. Treatment of the underlying disease is of central importance in controlling acute or chronic DIC associated with malignant diseases and chemotherapy should be started as soon as possible.
Lung dendritic cells facilitate extrapulmonary bacterial dissemination during pneumococcal pneumonia
(2013)
Streptococcus pneumoniae is a leading cause of bacterial pneumonia worldwide. Given the critical role of dendritic cells (DCs) in regulating and modulating the immune response to pathogens, we investigated here the role of DCs in S. pneumoniae lung infections. Using a well-established transgenic mouse line which allows the conditional transient depletion of DCs, we showed that ablation of DCs resulted in enhanced resistance to intranasal challenge with S. pneumoniae. DCs-depleted mice exhibited delayed bacterial systemic dissemination, significantly reduced bacterial loads in the infected organs and lower levels of serum inflammatory mediators than non-depleted animals. The increased resistance of DCs-depleted mice to S. pneumoniae was associated with a better capacity to restrict pneumococci extrapulmonary dissemination. Furthermore, we demonstrated that S. pneumoniae disseminated from the lungs into the regional lymph nodes in a cell-independent manner and that this direct way of dissemination was much more efficient in the presence of DCs. We also provide evidence that S. pneumoniae induces expression and activation of matrix metalloproteinase-9 (MMP-9) in cultured bone marrow-derived DCs. MMP-9 is a protease involved in the breakdown of extracellular matrix proteins and is critical for DC trafficking across extracellular matrix and basement membranes during the migration from the periphery to the lymph nodes. MMP-9 was also significantly up-regulated in the lungs of mice after intranasal infection with S. pneumoniae. Notably, the expression levels of MMP-9 in the infected lungs were significantly decreased after depletion of DCs suggesting the involvement of DCs in MMP-9 production during pneumococcal pneumonia. Thus, we propose that S. pneumoniae can exploit the DC-derived proteolysis to open tissue barriers thereby facilitating its own dissemination from the local site of infection.
Background/Aims: Only a small percentage of pathological gamblers utilizes professional treatment for gambling problems. Little is known about which social and gambling-related factors are associated with treatment utilization. The aim of this study was to look for factors associated with treatment utilization for pathological gambling. Methods: The study followed a sampling design with 3 different recruitment channels, namely (1) a general population-based telephone sample, (2) a gambling location sample and (3) a project telephone hotline. Pathological gambling was diagnosed in a telephone interview. Participants with pathological gambling (n = 395) received an in-depth clinical interview concerning treatment utilization, comorbid psychiatric disorders and social characteristics. Results: Variables associated with treatment were higher age [odds ratio (OR) 1.05, 95% confidence interval (CI) 1.03-1.08], an increased number of DSM-IV criteria for pathological gambling (OR 1.34, 95% CI 1.06-1.70), more adverse consequences from gambling (OR 1.10, 95% CI 1.03-1.16) and more social pressure from significant others (OR 1.17, 95% CI 1.07-1.27). Affective disorders were associated with treatment utilization in the univariate analysis (OR 1.81, 95% CI 1.19-2.73), but multivariate analysis showed that comorbid psychiatric disorders were not independently associated. Conclusion: These results indicate that individuals with more severe gambling problems utilize treatment at an older age when more adverse consequences have occurred. Further research should focus on proactive early interventions.
Hyperuricemia and its symptoms are becoming increasingly common worldwide. Elevated serum uric acid levels are caused by increased uric acid synthesis from food constituents and reduced renal excretion. Treatment in most cases involves reducing alcohol intake and consumption of meat and fish or treatment with pharmaceuticals. Another approach could be to reduce uric acid level in food, either during production or consumption. This work reports the production of recombinant urate oxidase by Arxula adeninivorans and its application to reduce uric acid in a food product. The A. adeninivorans urate oxidase amino acid sequence was found to be similar to urate oxidases from other fungi (61-65% identity). In media supplemented with adenine, hypoxanthine or uric acid, induction of the urate oxidase (AUOX) gene and intracellular accumulation of urate oxidase (Auoxp) was observed. The enzyme characteristics were analyzed from isolates of the wild-type strain A. adeninivorans LS3, as well as from those of transgenic strains expressing the AUOX gene under control of the strong constitutive TEF1 promoter or the inducible AYNI1 promoter. The enzyme showed high substrate specificity for uric acid, a broad temperature and pH range, high thermostability and the ability to reduce uric acid content in food.
Background: Primary angiitis of the central nervous system (PACNS) is a rare but serious condition. A fraction of patients suffering from PACNS concurrently exhibit pronounced cerebral amyloid angiopathy (CAA) which is characterized by deposits of amyloid-β (Aβ) in and around the walls of small and medium-sized arteries of the brain. PACNS with CAA has been identified as a distinct disease entity, termed Aβ-related angiitis (ABRA). Evidence points to an immune reaction to vessel wall Aβ as the trigger of vasculitis. Objective: To investigate whether the inflammatory response to Aβ has (1) any effect on the status of immune activation in the brain parenchyma and (2) leads to clearance of Aβ from brain parenchyma. Methods: We studied immune activation and Aβ load by quantitative immunohistochemical analysis in brain parenchyma adjacent to affected vessels in 11 ABRA patients and 10 matched CAA controls. Results: ABRA patients showed significantly increased immune activation and decreased Aβ loads in the brain parenchyma adjacent to affected vessels. Conclusion: Our results are in line with the hypothesis of ABRA being the result of an excessive immune response to Aβ and show that this can lead to enhanced clearance of Aβ from the brain parenchyma by immune-mediated mechanisms.
Background: This study aims to assess the role of ductoscopy for detecting intraductal anomalies in patients with nipple discharge in comparison to conventional tests and to find an effective combination of both approaches. Materials and Methods: Prior to duct excision, ductoscopy was performed in 97 women. Histologic and all other diagnostic results were compared. Sensitivity, specificity, and efficiency were calculated for all methods. These parameters were also calculated for all possible test combinations in 12 patients who had completed all tests. Results: Breast sonography reached the highest sensitivity (64.1%) and efficiency (64%); mammography had the highest specificity (100%). The sensitivity of ductoscopy was 53.2%, its specificity 60%, and its efficiency 55.1%. Among combinations of all methods, the combination ductoscopy + galactography was the most sensitive (80%). Mammography, magnetic resonance imaging, and ductoscopy were each 100% specific. Ductoscopy was the most efficient (75%) single method. Conclusion: Ductoscopy is a valuable test for diagnosing intraductal lesions in patients with nipple discharge. It is more efficient than conventional tests in patients undergoing all tests.
The objective of the present investigation was to examine the residual antimicrobial activity after a topical exposure of reconstructed human epidermis (RHE) to equimolar solutions of either chlorhexidine digluconate (CHG, 0.144% w/v) or octenidine dihydrochloride (OCT, 0.1% w/v) for 15 min. RHE-associated antiseptic agents were more effective on Staphylococcus aureus than on Pseudomonas aeruginosa. S. aureus was not detected after 24 h of contact, which demonstrated a microbicidal efficacy of greater than 5-log<sub>10</sub> reduction. In contrast, P. aeruginosa was reduced by approximately 2 log<sub>10</sub> at the same incubation time, which parallels the growth of the initial inoculum. This result could be interpreted either as a microbiostatic effect or as an adherence of P. aeruginosa to a low positively charged surface. Small amounts of CHG and OCT can penetrate the stratum corneum. Using these antiseptic agents, the viability of keratinocytes was reduced to 65-75% of that of the untreated RHE control following 24 h incubation in the presence of test microorganisms. With consideration of antimicrobial activity and cytotoxic effect, OCT corresponds better to a biocompatible antiseptic agent than CHG.
mikroRNAs (miRNAs oder miRs) sind kurze nicht-kodierende RNA-Moleküle, welche die Expression einer Vielzahl von Genen regulieren können. Das versetzt sie in die Lage, onkogene Zellsignalwege auf mehreren Ebenen fördern oder hemmen zu können. Die Rolle von miRNAs im Prostatakarzinom (PCa) wird zurzeit intensiv untersucht. Einen Beitrag dazu liefert diese Arbeit, welche die Rolle der miRNA miR-1 in PCa-Zellen charakterisiert. Untersucht wurde das Expressionsverhalten von Hitzeschock-Proteinen (HSPs) und Androgenrezeptor (AR) in PCa-spezifischen Zelllinien nach Überexpression bzw. Inhibition von miR-1. Des Weiteren wurde mit Hilfe einer Microarray-Untersuchung nach weiteren Zielen von miR-1-Regulation gesucht. Dabei erwies sich miR-1 als zentraler Faktor in einem regulatorischen Netzwerk, in dem sich HSP70, HSP90α, AR und TGF-β als Ziele miR-1-gesteuerter Suppression darstellten. miR-1 selbst unterliegt dabei ihrerseits der Regulation durch HSP27. Die Suppression von AR und HSPs hat letztlich anti-proliferative zelluläre Effekte zur Folge. Zum einen, weil HSP70 und AR pro-proliferativ wirken, zum anderen, da HSP70 und HSP90α den AR stabilisieren und dadurch seine pro-proliferative Funktion vermitteln. Auch das im Zuge einer Mircoarray-Untersuchung identifizierte miR-1-Ziel TGF-β wird durch diese miRNA negativ reguliert, was ebenfalls anti-proliferative Wirkung auf PCa-Zellen hat. Zudem wird TGF-β mit Metastasierungs-Prozessen in Verbindung gebracht, auf die miR-1, über die Suppression von TGF-β, möglicherweise regulatorisch einwirken kann. Die in dieser Arbeit gezeigte Herabregulation von miR-1 in menschlichem PCa-Gewebe bestätigt die in vitro Ergebnisse und lässt die Schlussfolgerung zu, dass miR-1 die Funktion eines Tumorsuppressors im PCa besitzt, welcher im Zuge der Malignisierung des Karzinoms, im Sinne eines Resistenz-Mechanismus, herunterreguliert wird. Damit stellt die miR-1 ein mögliches Ziel zukünftiger pharmakologischer Intervention bei der Therapie des PCa dar.
Das Ziel dieser Arbeit ist es auf Basis einer großen populationsbasierten Kohorte im Rahmen der SHIP-Studie (Study of Health of Pommerania) die mögliche Assoziation zwischen der Serum PRL-Konzentration mit dem MetS und dem T2DM aufzuzeigen. Dieser Sachverhalt wurde bereits in früheren ausgewählten Studien mit kleineren Kohorten untersucht. In unserer Studie wurden dazu die Daten von 3,993 Individuen (2,027 Frauen) in einem Alter von 20-79 Jahren aus der populationsbasierten SHIP-Studie verwendet. Die Assoziation zwischen PRL-Konzentrationen und MetS sowie dem T2DM wurden sowohl im Queer- als auch im Längsschnitt mittels alters- und multivariabel-adjustierten Poisson-Regressionsmodellen untersucht. PRL wurde log-transformiert und als kontinuierliche (per Anstieg der Standartabweichung (SD)) oder kategoriale (geschlechtsspezifische Quartil) Einflussvariable, getrennt nach Männern und Frauen, dargestellt. Die Querschnittsanalyse zeigte eine inverse Assoziation zwischen niedrigen PRL Konzentrationen und einem prävalenten T2DM Risiko sowohl in Männern als auch in Frauen nach multivariabler Adjustierung (Männer: Q1 vs. Q4: relatives Risiko (RR), 1,55; 95% Konfidenzintervall (CI), 1.13 – 2.14; Frauen: Q1 vs. Q4: RR, 1.70; 95% CI, 1.10 – 2.62). Gleichermaßen wurde höhere PRL Konzentrationen mit signifikant niedrigerem T2DM Risiko assoziiert (RR pro SD Anstieg in log-transformierten PRL: 0,83, 95% CI, 0,72-0,95 bei Männern und 0,84, 95% CI, 0,71 bis 0,98 bei Frauen). Die inverse Assoziation zwischen PRL und dem MetS konnte nach der multivariablen Adjustierung nicht beibehalten werden. In der Längsschnittanalyse konnte die Assoziation zwischen PRL und inzidentem MetS oder T2DM nicht aufrechterhalten werden. Zusammenfassend ist dies die erste große populationsbasierte Studie, welche im Querschnitt über eine inverse Assoziation zwischen PRL und prävalentem T2DM in beiden Geschlechtern berichten kann. Jedoch deutet die fehlende longitudinale Assoziation darauf hin, dass PRL keine kausale Rolle als Risikofaktor für einen inzidenten T2DM oder MetS darstellt.