Search

Oncogenic types of human papillomaviruses (HPVs) are major human carcinogens. The viral E6/E7 oncogenes maintain the malignant growth of HPV-positive cancer cells. Targeted E6/E7 inhibition results in efficient induction of cellular senescence, which could be exploited for therapeutic purposes. Here we show that viral E6/E7 expression is strongly downregulated by Metformin in HPV-positive cervical cancer and head and neck cancer cells, both at the transcript and protein level. Metformin-induced E6/E7 repression is glucose and PI3K-dependent but-other than E6/E7 repression under hypoxia-AKT-independent. Proteome analyses reveal that Metformin-induced HPV oncogene repression is linked to the downregulation of cellular factors associated with E6/E7 expression in HPV-positive cancer biopsies. Notably, despite efficient E6/E7 repression, Metformin induces only a reversible proliferative stop in HPV-positive cancer cells and enables them to evade senescence. Metformin also efficiently blocks senescence induction in HPV-positive cancer cells in response to targeted E6/E7 inhibition by RNA interference. Moreover, Metformin treatment enables HPV-positive cancer cells to escape from chemotherapy-induced senescence. These findings uncover profound effects of Metformin on the virus/host cell interactions and the phenotype of HPV-positive cancer cells with implications for therapy-induced senescence, for attempts to repurpose Metformin as an anticancer agent and for the development of E6/E7-inhibitory therapeutic strategies.

Molluscum contagiosum (MC) is an infectious disease that occurs only in humans with a tropism that is narrowly restricted to the outermost epidermal layer of the skin. Molluscum contagiosum virus (MCV) is the causative agent of MC which produces skin lesions that can persist for months to several years. MCV is efficiently transmitted by direct physical contact or by indirect contact with fomites. MC is most prevalent in children and immune compromised patients. The failure to develop a drug that targets MCV replication has been hampered for decades by the inability to propagate MCV in cell culture. To address this dilemma, we recently engineered a surrogate poxvirus expressing the MCV processivity factor (mD4) as the drug target. The mD4 protein is essential for viral replication by keeping the viral polymerase tethered to the DNA template. In this study we have designed and synthesized a lead compound (7269) that is able to prevent mD4 dependent processive DNA synthesis in vitro (IC

ZusammenfassungEin 44-jähriger Patient hatte anhaltendes Fieber und eine deutliche Erhöhung des C-reaktiven Proteins (CRP) nach einer Infektion mit dem severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) Bei zunehmender Sinustachykardie erfolgte die Bestimmung der Schilddrüsenhormone, welche eine Hyperthyreose zeigten. Sonographisch fand sich eine Vergrößerung der Schilddrüse mit unscharf begrenzten echoarmen Arealen. Wir stellten die Diagnose einer SARS-CoV-2-Infektions-assoziierten subakuten Thyreoiditis und leiteten eine Therapie mit Prednisolon ein. Diese Therapie führte kurzfristig zu einer Besserung des klinischen Zustands und zu einer vollständigen Ausheilung binnen drei Monaten.

A 44-year-old man had persistent fever following a severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infection. Because of progressive sinus tachycardia, thyroid hormones were measured which showed hyperthyroidism. Thyroid sonography revealed enlargement of the thyroid gland with hypoechoic areas with blurred margins. We diagnosed subacute granulomatous thyroiditis associated with SARS-CoV‑2 infection and initiated therapy with prednisolone. This therapy resulted in rapid improvement of the patient's clinical condition and complete remission after three months.Ein 44-jähriger Patient hatte anhaltendes Fieber und eine deutliche Erhöhung des C-reaktiven Proteins (CRP) nach einer Infektion mit dem severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) Bei zunehmender Sinustachykardie erfolgte die Bestimmung der Schilddrüsenhormone, welche eine Hyperthyreose zeigten. Sonographisch fand sich eine Vergrößerung der Schilddrüse mit unscharf begrenzten echoarmen Arealen. Wir stellten die Diagnose einer SARS-CoV-2-Infektions-assoziierten subakuten Thyreoiditis und leiteten eine Therapie mit Prednisolon ein. Diese Therapie führte kurzfristig zu einer Besserung des klinischen Zustands und zu einer vollständigen Ausheilung binnen drei Monaten.

The smallpox virus (variola) remains a bioterrorism threat since a majority of the human population has never been vaccinated. In the event of an outbreak, at least two drugs against different targets of variola are critical to circumvent potential viral mutants that acquire resistance. Vaccinia virus (VACV) is the model virus used in the laboratory for studying smallpox. The VACV processivity factor D4 is an ideal therapeutic target since it is both essential and specific for poxvirus replication. Recently, we identified a tripeptide (Gly-Phe-Ile) motif at the C-terminus of D4 that is conserved among poxviruses and is necessary for maintaining protein function. In the current work, a virtual screening for small molecule mimics of the tripeptide identified a thiophene lead that effectively inhibited VACV, cowpox virus, and rabbitpox virus in cell culture (EC50 = 8.4–19.7 μM) and blocked in vitro processive DNA synthesis (IC50 = 13.4 μM). Compound-binding to D4 was demonstrated through various biophysical methods and a dose-dependent retardation of the proteolysis of D4 proteins. This study highlights an inhibitor design strategy that exploits a susceptible region of the protein and identifies a novel scaffold for a broad-spectrum poxvirus inhibitor.

Background In order to guide therapy in Ulcerative Colitis (UC), repeated determination of intestinal inflammatory activity is essential. Endoscopy is the standard procedure to assess inflammation in UC. However innovative methods for non-invasive, uncomplicated and risk free estimation of inflammatory activity are needed as bowel preparation, patients discomfort and risk of procedural complications limit the (frequent) use of colonoscopy. Multispectral optoacoustic tomography (MSOT) is a promising new method to measure inflammation in UC. Using short and harmless impulses of NIR-lasers, it allows for determination of a specific hemoglobin-signal in the bowel-wall and therefore inflammatory activity in affected bowel segments. However, its informative value in UC has not been evaluated so far. Methods In 34 patients with confirmed UC, clinical activity parameters (e.g. clinical Mayo-Subscore, B-mode-sonography, C-reactive protein, white blood count) were collected and MSOT of the sigmoid was performed within 2 weeks before/after endoscopy. For MSOT, a commercially available clinical MSOT-system (Acuity Echo, iThera Medical, Munich) was used with sequential analysis of collected data on an external desktop PC. Finally, clinical data, ultrasound findings (Limberg) and MSOT-parameters (single wavelenghts 760 nm, 800 nm, 900 nm; multispectral signals hb, hbO2, hbT) were correlated with endoscopic findings (Mayo endoscopic Subscore, MES). Results We found strong and significant correlation between MES and MSOT parameters 800 nm (Spearman r = 0,6599; p < 0,0001) and HbO2 (Spearman r = 0,6695; p < 0,0001), superior to sonographic evaluation of the inflammatory activity in affected bowel segments (Spearman r = 0,4914; p = 0,0023). Simultaneously these MSOT parameters demonstrated excellent sensitivity and specifity in distinguishing moderately to highly active (MES 2,3) from inactive and mild disease (MES 0,1) (800nm: AUROC 0,9063 (p < 0,0001); sensitivity = 93,75 %, specificity = 88,89 %; HbO2: AUROC 0,9063 (p < 0,0001); sensitivity = 100 %, specificity = 88,89 %). Conclusion MSOT is a promising approach to non-invasively assess intestinal inflammation in UC and therefore monitor anti-inflammatory therapy in these patients. Further studies are required to validate these findings.

Zymoseptoria tritici is the causal agent of septoria tritici blotch (STB), a foliar wheat disease important worldwide. Succinate dehydrogenase inhibitors (SDHIs) have been used in cereals for effective control of STB for several years, but resistance towards SDHIs has been reported in several phytopathogenic fungi. Resistance mechanisms are target-site mutations in the genes coding for subunits B, C and D of the succinate dehydrogenase (SDH) enzyme. Previous monitoring data in Europe indicated the presence of single isolates of Z. tritici with reduced SDHI sensitivity. These isolates carried mutations leading to amino acid exchanges: C-T79N, C-W80S in 2012; C-N86S in 2013; B-N225T and C-T79N in 2014; and C-V166M, B-T268I, C-N86S, C-T79N and C-H152R in 2015. The current study provides results from microtitre and greenhouse experiments to give an insight into the impact of different mutations in field isolates on various SDHIs. In microtitre tests, the highest EC50 values for all tested SDHIs were obtained with mutants carrying C-H152R. Curative greenhouse tests with various SDHIs confirmed the findings of microtitre tests that isolates with C-H152R are, in general, controlled with lower efficacy than isolates carrying B-T268I, C-T79N and C-N86S. SDHI-resistant isolates of Z. tritici found in the field were shown to have cross-resistance towards all SDHIs tested. So far, SDHI-resistant isolates of Z. tritici have been found in low frequencies in Europe. Therefore, FRAC recommendations for resistance management in cereals, including a limited number of applications, alternation and combination with other MOAs, should be followed to prolong SDHI field efficacy. This article is protected by copyright. All rights reserved.

Rare truncating BRCA2 K3326X (rs11571833) and pathogenic CHEK2 I157T (rs17879961) variants have previously been implicated in familial pancreatic ductal adenocarcinoma (PDAC), but not in sporadic cases. The effect of both mutations in important DNA repair genes on sporadic PDAC risk may shed light on the genetic architecture of this disease. Both mutations were genotyped in germline DNA from 2,935 sporadic PDAC cases and 5,626 control subjects within the PANcreatic Disease ReseArch (PANDoRA) consortium. Risk estimates were evaluated using multivariate unconditional logistic regression with adjustment for possible confounders such as sex, age and country of origin. Statistical analyses were two-sided with p values

Cash flow planning for owners of closely held businesses is an important component to the ultimate success and continuation of that business. This article presents a number of important planning issues that require careful consideration concerning the adequacy of cash flow generation. Looking ahead and planning for the smooth transition of a business to new management is necessary for the financial well-being of a family. © 2015 Wiley Periodicals, Inc.

This article considers the many factors that must go into developing a transfer price to be used as products and services move between entities and divisions of multinational enterprises (MNEs). The discussion focuses on two different economic theories: resource-based value and transaction cost economics. These two theories are integrated to better understand the multitude of factors that must be considered in developing a transfer price. The use of a well-thought-out transfer price has a positive effect on the value of a firm. © 2015 Wiley Periodicals, Inc.

Protein disulfide isomerase (PDI, PDIA1) is an emerging therapeutic target in oncology. PDI inhibitors have demonstrated a unique propensity to selectively induce apoptosis in cancer cells and overcome resistance to existing therapies, although drug candidates have not yet progressed to the stage of clinical development. We recently reported the discovery of lead indene compound E64FC26 as a potent pan-PDI inhibitor that enhances the cytotoxic effects of proteasome inhibitors in panels of Multiple Myeloma (MM) cells and MM mouse models. An extensive medicinal chemistry program has led to the generation of a diverse library of indene-containing molecules with varying degrees of proteasome inhibitor potentiating activity. These compounds were generated by a novel nucleophilic aromatic ring cyclization and dehydration reaction from the precursor ketones. The results provide detailed structure activity relationships (SAR) around this indene pharmacophore and show a high degree of correlation between potency of PDI inhibition and bortezomib (Btz) potentiation in MM cells. Inhibition of PDI leads to ER and oxidative stress characterized by the accumulation of misfolded poly-ubiquitinated proteins and the induction of UPR biomarkers ATF4, CHOP, and Nrf2. This work characterizes the synthesis and SAR of a new chemical class and further validates PDI as a therapeutic target in MM as a single agent and in combination with proteasome inhibitors.

Bei klinisch vorhandenen Muskelschmerzen, einer massiven Erhohung der CK im Serum und Myoglobulinurie liegt eine Rhabdomyolyse vor. Ursachen konnen neben Trauma, myotoxischen Medikamenten, Alkohol-, Drogenkonsum und Infektionen auch endokrine Stoffwechselstorungen oder Autoimmunerkrankungen sein. Auftretende Komplikationen sind beispielsweise akutes Nierenversagen und Kompartmentsyndrom. Das fruhzeitige Erkennen der Ursache und deren Behebung und eine adaquate symptomatische Therapie verhindern schwerwiegende Komplikationen. Wir prasentieren den klinischen Fall einer 44-jahrigen Patientin, die sich mit dem Leitsymptom Myalgie vorstellte. Bei CK-Werten bis 8 830 IU/l. Begleitend zeigte sich ein Perikarderguss, ein Myxodem der Stimmlippen und im Verlauf trat eine Verschlechterung der Nierenfunk­tion auf. Unter L-Thyroxin Substitution und Volumensubstitution waren die Beschwerden rucklaufig. Als Ursache diagnostizierten wir eine Hashimoto-Thyreoiditis. Eine isolierte Hypothyreose als Ursache einer Rhabdomyolyse ist selten. In der Literatur finden sich haufig auslosende Kofaktoren in der Anamnese, vor allem eine Therapie mit HMG-CoA-Reduktase-Hemmern. Bei Myalgien und CK-Erhohung als Leitsymptom muss als Ursache neben anderen Auslosern an eine Hashimoto-Thyreoiditis gedacht werden. Durch Hormonsubstitution und adaquate symptomatische Therapie konnen Komplikationen vermieden werden.

The approval of microbubbles with the inert gas sulfur hexafluoride (SF6) and a palmitic acid shell (SonoVue(®), Bracco Geneva, CH) for the diagnostic imaging of liver tumors in adults and children by the FDA in the United States represents a milestone for contrast-enhanced ultrasound (CEUS).This warrants a look back at the history of the development of CEUS. The first publications based on echocardiographic observations of right ventricular contrast phenomena caused by tiny air bubbles following i. v. injection of indocyanine green appeared around 1970 1 2 3. A longer period of sporadic publications but no real progress then followed since, in contrast to X-ray methods, ultrasound works quite well without a contrast agent.It is noteworthy that the foundations for further development were primarily laid in Europe. The development and approval (1991) of the contrast agent Echovist(®) by a German contrast manufacturer for echocardiography unsuitable for passing through lungcapillaries 4 5 resulted in the first extracardiac indications, e. g. for detecting retrovesical reflux and tubal patency, in the mid-1980 s 6 7 8. The sensitivity of color Doppler was not able to compensate for the lack of an ultrasound contrast agent compared to CT with its obligatory contrast administration.Studies of SHU 508 - microbubbles of air moderately stabilized with galactose and palmitic acid - began in 1990 9 10 11 12 13 14 15 and the contrast agent was then introduced in 1995 in Germany as Levovist(®). The most important publications by Blomley, Cosgrove, Leen, and Albrecht are named here on a representative basis 16 17 18 19 20.SHU 508 along with other US contrast agents provided impressive proof of the superiority of CEUS for the diagnosis of liver metastases. However, practical application remained complicated and required skill and technical know-how because of a lack of suitable software on US units 21 22 23 24 25. The monograph regarding the use of contrast agent in the liver by Wermke and Gaßmann is impressive but unfortunately only available in German 26. In addition to being applied in the heart and the liver, CEUS was first used in transcranial applications 27 and in vessels 28, the kidneys 29, and the breast 30. Measurements at transit times were also of particular interest 31. It was difficult to convince ultrasound device manufacturers of the need to adapt US units to US contrast agents and not vice versa.The breakthrough came with low MI phase contrast inversion and the introduction of SonoVue(®) in many European countries in 2001. This more stable US contrast agent is easy to use and is becoming indispensable in diagnostic imaging of the liver 32 33 34 35 36 37 38 39 40. Studies have shown its excellent tolerability 41 and diagnostic reliability comparable to that of MDCT and MRI in the liver 42 43. Today it would be unimaginable to diagnose liver tumors without CEUS 44. This also applies to very small lesions 45 46.EFSUMB published the first CEUS guidelines in 2004 47 which have since been reissued and divided into hepatic 48 and extrahepatic applications 49. The first recommendations regarding quantitative assessment have also been published 50.The increasing scientific interest in CEUS is evident based on the greater number of PubMed hits for Echovist(®) (ca. 130), Levovist(®) (ca. 500) and SonoVue(®) (ca. 1500) as well as on the fact that publications regarding CEUS comprise almost 20 % of UiM/EJU articles in the last 10 years. The number of CEUS articles in UiM/EJU continues to be high 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75.In the clinical reality, CEUS has been able to become established alongside CT and MRI according to the saying "better is the enemy of good" 76 as the method of choice after B-mode ultrasound in the evaluation of liver tumor malignancy in Germany, where the technically challenging method is promoted. In the case of unclear CT and MRI findings, CEUS performed by an experienced examiner/clinician often provides the solution, particularly in the case of small lesions, and is the last resort before US-guided biopsy 45 46. However, there is a lack of competent CEUS examiners and Germany continues to be the world champion of X-ray examinations with no noticeable reverse trend. In almost every doctor's office and hospital, ultrasound costs are by far not fully covered, resulting in an extremely high frequency of CT use with CT being available to everyone regardless of insurance status.The USA is now in the starting position for CEUS. It will be exciting to see how the method will develop there. The FDA's decision to approve sulfur hexafluoride (Lumason(®) = SonoVue(®)) should be considered against the background of the radiation exposure caused by CT examinations and the fact that MRI using gadolinium-containing contrast agents is no longer considered noninvasive because of nephrogenic systemic fibrosis (NSF) and the accumulation of the agent in the cerebrum. An essential point of the campaign regarding the avoidance of diagnostic radiation exposure triggered in the USA by the publications of Brenner et al. 77 78 was that the agent was approved for use in the liver even for children 79 80 - still off label in Europe - without additional comprehensive studies due to the available scientific results and the very low side effects profile of Lumason(®) (= SonoVue(®)). It is admittedly unclear why other indications (except the heart which has been approved since 2014) are excluded even though the microbubbles as a pure blood pool contrast agent can be diagnostically used in the entire vascular system and bed of all organs. To our knowledge, there is no such restriction on the approval of X-ray contrast agents.Like echocardiography and emergency ultrasound, CEUS began in Europe but will probably only establish its final diagnostic value as a "reimport".This is a major opportunity to permanently define the role of Ultrasound as a highly valuable, patient-centered imaging method in the German health care system.This may prompt some of our international readers to reflect upon the role of CEUS in their own countries.

Hintergrund: Pfortaderthrombosen treten mit einer Pravalenz von 1% bei nicht-vorgeschadigter Leber auf. Ursachlich ist eine erhohte systemische oder lokale Thrombophilieneigung bei myeloproliferativen Erkrankungen, Malignomen, Gerinnungsstorungen oder intraabdominellen Entzundungen, aber auch systemischen Virusinfekten mit Begleithepatitis. Therapie der Wahl ist die sofortige Vollantikoagulation uber 3 – 6 Monate (initial mit niedermolekularem Heparin, im Verlauf mit Vitamin-K-Antagonisten) [3; 4]. In der Literatur finden sich Fallberichte uber Pfortaderthrombosen bei Immunkompetenten bei CMV-Primarinfektionen [2; 4; 5]. Bislang nicht beschrieben sind Assoziationen mit anderen Viren der Herpesgruppe. Wir prasentieren den Fall mit bildgebenden Befunden einer partiellen Pfortaderthrombose nach EBV-Infektion mit diskreter Begleithepatitis bei einem jungen Mann ohne relevante Vorerkrankungen mit negativem Thrombophilie-Screening. Die orale Antikoagulation erfolgte nach initialer Heparinisierung mit Rivaroxaban, auch vor dem Hintergrund, dass Warfarin-Nekrosen bei EBV-Infektionen beschrieben sind [1]. Nach 4 Wochen zeigte sich die Pfortaderthrombose vollstandig aufgelost; der Patient ist seither beschwerdefrei. Fallbericht: Ein 27-jahriger, deutschstammiger Patient stellte sich mit seit einer Woche bestehender Abgeschlagenheit in unserer internistischen Notaufnahme vor. Der klinische Untersuchungsbefund war unauffallig. 4 Monate zuvor hatte er eine EBV-Infektion mit leichter Leberbeteiligung durchgemacht. Nach zwischenzeitlicher Normalisierung der Laborwerte war nun eine leicht erhohte ALT von 63,2 U/L (Referenzbereich: 10 – 50 U/L) bei normwertiger AST und GGT aufgefallen. Sonographisch zeigte sich eine komplette Pfortaderthrombose des linken Pfortaderastes und partielle Thrombose des rechten Pfortaderastes bei unauffalligem Leberparenchymbefund ohne fokale Lasionen. Die ubrige Abdomensonografie inklusive Darmsonografie war unauffallig. Laborchemisch zeigten sich lediglich ein minimal erhohtes CRP (12,3 mg/L; Referenzbereich: < 5 mg/L), eine LDH von 270 U/L (< 250 U/L) und eine ALT von 61 U/L (< 50 U/L). Der Patient wurde nach initialer Heparinisierung mit Rivaroxaban 2 × 15 mg therapeutisch antikoaguliert. Ein umfangreiches Thrombophilie-Screening blieb trotz familienanamnestisch erhohter Thrombophilieneigung unauffallig. An Vorerkrankungen waren eine Amoxicillin-Allergie und ein laparoskopischer Leistenhernienverschluss vor 7 Monaten bekannt. Eine Kontrollsonografie nach 4 Wochen oraler Antikoagulation zeigte eine vollstandige Rekanalisierung des Pfortadersystems. Samtliche Leberwerte waren normalisiert. Eine weitere Verlaufskontrolle nach 16 Wochen war ebenfalls unauffallig; die orale Antikoagulation wurde daraufhin beendet. Schlussfolgerungen: Der vorgestellte Fall einer akuten Pfortaderthrombose nach EBV-Infektion mit diskreter Begleithepatitis zeigt Parallelen zu den in der Literatur beschriebenen Fallberichten bei CMV-Primarinfektionen immunkompetenter Patienten. Pathogenetisch werden direkte Viruseffekte auf das Gefasendothel ebenso diskutiert wie ein Zusammenhang der Antikorper-Titer mit prokoagulatorischen Faktoren und die Begleithepatitis als lokaler Trigger [2; 5]. Meist zeigen sich nur minimal erhohte Laborparameter (AST, ALT, LDH und D-Dimere); Bauchschmerzen konnen oft fehlen. Auch unser Patient war klinisch asymptomatisch. Aufgrund des diskreten Anstiegs der Transaminasen erfolgte eine Lebersonografie, die den Befund der Pfortaderthrombose ergab. Der vorliegende Fall zeigt, dass auch im Rahmen anderer Virusinfekte, insbesondere bei Vorliegen einer Begleithepatitis, an die Moglichkeit einer Pfortaderthrombose gedacht werden sollte. Rivaroxaban scheint dabei eine therapeutische Alterative zu Phenprocoumon darzustellen. Auch bei Vorliegen einer plausiblen lokalen Ursache der Pfortaderthrombose (serologischer Nachweis einer Virusinfektion mit Begleithepatitis) sollte eine zusatzliche, bislang unbekannte Thrombophilieneigung ausgeschlossen werden. Literatur: 1. Franson TR, Rose HD, Spivey MR, et al. Late-onset, warfarin-caused necrosis occurring in a patient with infectious mononucleosis. Arch Dermatol. 1984 Jul;120(7):927 – 31. 2. Gueddi S, Righini M, Mezger N, et al. Portal vein thrombosis following a primary cytomegalovirus infection in an immunocompetent adult. Thromb Haemost. 2006 Jan;95(1):199 – 201. 3. Plessier A, Rautou PE, Valla DC. Management of hepatic vascular diseases. J Hepatol. 2012;56 Suppl 1:S25 – 38. 4. Primignani M. Portal vein thrombosis, revisited. Dig Liver Dis. 2010 Mar;42(3):163 – 70. 5. Squizzato A, Ageno W, Cattaneo A, et al. A case report and literature review of portal vein thrombosis associated with cytomegalovirus infection in immunocompetent patients. Clin Infect Dis. 2007 Jan 15;44(2):e13 – 6.

PURPOSE: Postsurgical evaluation of bone microcirculation in osseous and osseocutaneus free flaps by contrast enhanced ultrasound (CEUS) with time intensity curve (TIC) analysis and [18F]-positron emission tomography/computed tomography (Fluoride-PET/CT). PATIENTS AND METHODS: 10 patients with osseous or osseocutaneus microvascular flaps were evaluated with CEUS. Ultrasound was carried out by an experienced examiner with a high resolution linear probe (6-9 MHz, LOGIQ E9, GE) after intravenous bolus injection of 2.4 ml SonoVue® (BRACCO, Germany). Time intensity curves (TIC) in selected regions of interest (ROI) were analyzed and compared with the evaluation of [18F]-positron emission tomography/computed tomography and the clinical course for at least 2 month. RESULTS: 9/10 patients showed evidence for soft tissue and osseous microcirculation of the transplants in CEUS correlating with the clinical course. The soft tissue parts of the transplant showed a mean value of 84% (range 51-98%) and the bone parts a mean value of 39% (range 11-75%) for the Area under the curve (AuC) compared with the AuC for the anastomotic vessel region. Mean values for time to peak (TTP) were 27,1 sec (range 8.7-52.0 sec) for the anastomotic vessels, 29.3 sec (range 7.9-62.0 sec) for the soft tissue of and 32.0 sec (range 7.4-69.0 sec) for the transplant bone. In 1/10 patients flap failure occurred, after denudation the bone was left as an avascular transplant. AuC showed a mean value of 0.5 % for the bone region compared with the vessel region. Fluoride-PET/CT assessed bone vitality in 7 patients as "good" in 1 patient as "uncertain" and in 1 patient as "poor". CEUS assessment was corresponding with Fluoride-PET/CT in 4 patients, clinical assessment in 6 patients. CONCLUSION: Fluoride-PET/CT is a valuable tool to make an indirect statement about the perfusion of the transplanted bone and was used as control in this study. CEUS is a new and promising method for the evaluation of microcirculation of buried free microvascular bone grafts and the osseous part of osseocutaneous flaps and may be used for a steady monitoring in the first postoperative days.