Complementary analysis of the hard and soft protein corona: sample preparation critically effects corona composition.

Here we demonstrate how a complementary analysis of nanocapsule-protein interactions with and without application media allows gaining insights into the so called hard and soft protein corona. We have investigated how both human plasma and individual proteins (human serum albumin (HSA), apolipoprotein A-I (ApoA-I)) adsorb and interact with hydroxyethyl starch (HES) nanocapsules possessing different functionalities. To analyse the hard protein corona we used sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and a protein quantitation assay. No significant differences were observed with regards to the hard protein corona. For analysis of the soft protein corona we characterized the nanocapsule-protein interaction with isothermal titration calorimetry (ITC) and dynamic light scattering (DLS). DLS and ITC measurements revealed that a high amount of plasma proteins were adsorbed onto the capsules' surface. Although HSA was not detected in the hard protein corona, ITC measurements indicated the adsorption of an HSA amount similar to plasma with a low binding affinity and reaction heat. In contrast, only small amounts of ApoA-I protein adsorb to the capsules with high binding affinities. Through a comparison of these methods we have identified ApoA-I to be a component of the hard protein corona and HSA as a component of the soft corona. We demonstrate a pronounced difference in the protein corona observed depending on the type of characterization technique applied. As the biological identity of a particle is given by the protein corona it is crucial to use complementary characterization techniques to analyse different aspects of the protein corona.

Loss of PIDD limits NF-κB activation and cytokine production but not cell survival or transformation after DNA damage.

Activation of NF-κB (nuclear factor of kappa light chain gene enhancer in B cells) in response to DNA damage is considered to contribute to repair of genetic lesions, increased cell survival and cytokine release. The molecular mechanisms orchestrating this cytoplasmic event involve core components of the nuclear DNA damage response machinery, including ATM-kinase (ataxia telangiectasia mutated kinase) and PARP-1 (poly (ADP-ribose) polymerase 1). The physiological consequences of defective NF-κB activation in this context, however, remain poorly investigated. Here we report on the role of the 'p53-induced protein with a death domain', PIDD, which appears rate limiting in this process, as is PARP-1. Despite impaired NF-κB activation, DNA damage did not increase cell death or reduce clonal survival of various cell types lacking PIDD, such as mouse embryonic fibroblasts or stem and progenitor cells of the hematopoietic system. Furthermore, lymphomagenesis induced by γ-irradiation (IR) was unaffected by deficiency for PIDD or PARP-1, indicating that loss of DNA damage-triggered NF-κB signalling does not affect IR-driven tumorigenesis. However, loss of either gene compromised cytokine release after acute IR injury. Hence, we propose that NF-κB's most notable function after DNA damage in primary cells is related to the release of cytokines, thereby contributing to sterile inflammation.

[Traumatic optic neuropathy: a review of the literature in the light of personal experiences]. / Die traumatische Optikusneuropathie: Aktuelle Literaturübersicht im Spiegel eigener Erfahrungen.

BACKGROUND: Therapy of traumatic optic neuropathy (TON) is still discussed controversially. Studies of medical treatment and surgical decompression of the nerve could not find any correlation between therapy and result. Today's knowledge of the treatment in TON is to be analyzed by the latest results in the literature, supplemented by personal experiences with our own patients, who underwent a combination of corticosteroids and surgical decompression. METHODS: The study group consisted of 9 patients at the age of 13-58 years. 8 patients suffered from a cranial trauma, 1 patient had sinus surgery, which resulted in an indirect damage of the optic nerve. Pretherapeutically, 5 patients had residual vision, 4 patients were blind. A fracture line through the optic canal in the CT-scan was seen in 6 cases. Decompression was performed within 24 hours in 3 cases; in the worst 3 cases it took up to 8 days. In 8 patients the intervention was performed via an endonasal, microscopic-endoscopic approach, once it was done transfacially. Simultaneously, high-dose corticosteroids were administered. RESULTS: All patients with a residual vision before therapy showed an improvement of their visual acuity: In the best case visual acuity changed from perception of light to 0.8. All patients with posttraumatic blindness remained blind after therapy. CONCLUSION: A surgical decompression may be considered in patients with residual vision. Referring to the latest data in the literature endonasal, microscopic-endoscopic decompression is then to be combined with simultaneous application of high-dose corticosteroids. In our opinion, a mere wait-and-see strategy completely without any treatment can hardly be recommended.

[Multifocal manifestation of Rosai-Dorfman disease. A rare case of purely extranodal sinus histiocytosis in the elderly]. / Multifokale Manifestation einer Rosai-Dorfman-Erkrankung. Seltene, rein extranodale Sinushistiozytose im Erwachsenenalter.

Rosai-Dorfman disease belongs to the group of childhood histiocytoses and was initially described as sinus histiocytosis with massive lymphadenopathy. Its rare purely extranodal manifestation is primarily found in the head and neck region. An atypical primary manifestation in an elderly patient with multifocal extranodal disease is described, and this pathological entity is reviewed. Specific difficulties concerning differential diagnostic aspects as well as individually appropriate treatment strategies are discussed.

[Atypical gout tophi for differential diagnosis of head and neck tumors : case report and review of the literature]. / Atypische Gichttophi als Differenzialdiagnose von HNO-Tumoren : Fallbericht und Literaturübersicht.

Gout is a mostly hereditary metabolic disease and is considered a disease of affluence. The disease is promoted by a purine-rich diet and shows an intermittent course of inflammatory joint manifestations and periods free of symptoms. The pathognomonic sign of the disease is an acute and very painful monarthritis with typical local deposits of uric acid, so-called gout tophi. No or inadequate treatment leads to the chronic form of gouty arthritis characterized more by joint destruction than by persistent pain. In head and neck gout tophi are seen as nodular lesions along the outer helical edges of the auricle. A case report of gout manifestation in the infratemporal fossa, deriving from the temporomandibular joint, with arrosion of the bony skull base demonstrates gout as a relevant disease for the ENT clinician. Potential diagnostic difficulties as well as recommendations for a therapeutic regimen of gouty lesions in such critical localizations will be reviewed.

[Short and long term results of endolymphatic sac surgery: a patient-questionnaire based study]. / Die Saccotomie in der Behandlung des M. Ménière: Beurteilung der kurz-, mittel- und langfristigen Ergebnisse anhand eines Patientenfragebogens.

INTRODUCTION: The endolymphatic sac surgery for the treatment of Meniere's disease has been described since the 1920s. The success rate of this technique in terms of vertigo control has been reported to be 50-80%. However, the value of this treatment method remained controversial. Furthermore, the reliable identification of the endolymphatic sac intraoperatively can be challenging in some cases. This study examines the short-, middle- and long-term results in a larger cohort of patients. MATERIALS AND METHODS: In 74 patients, vertigo control, tinnitus and degree of satisfaction was evaluated by means of a questionnaire retrospectively. Additionally, the diagnostic value of the electrocochleography (EcochG) was determined. RESULTS: The overall vertigo control rate was more than 70% in patients followed up for two years and has reached 81% in patients followed up for more than two years. Hearing preservation rate was 61%. Tinnitus has disappeared in 11% and improved in 23% of the patients. In 47% of the patients it was unchanged and in 19% worsened. The difference in EcochG results pre- versus postoperative was highly significant. CONCLUSIONS: ELSS is a useful tool in the management of Ménière's disease, in particular in patients that do not benefit sufficiently from conservative therapy.

[Surgical complications of paranasal sinus surgery. Topographical classification and therapeutic management]. / Komplikationen der Nasennebenhöhlen-Chirurgie. Topographische Einteilung und therapeutisches Management.

Surgery of the paranasal sinuses is one of the most frequently performed surgical interventions in otorhinolaryngology today. The potential for surgical complications with damage to paranasal structures in the close vicinity (orbit, endocranium, vital arteries) is still significant despite very elaborate, minimally invasive endoscopic and microscopic techniques. Reviewing the wide range of surgical complications of paranasal sinus surgery in a systematic and topographical fashion, elucidated by typical clinical case reports, we discuss the therapeutic management of such surgical sequelae. In addition, medicolegal aspects of informed patient consent as well as general recommendations to possibly avoid such complications are given.

[Surgical therapy in Menière's disease. Historical development and today's state of the art]. / Die chirurgische Therapie des M. Menière. Historische Entwicklung und heutiger Stand.

After P. Menière's first description of the typical symptoms in 1861 it took more than 40 years before the first otosurgical procedures were performed to cure Menière's disease. Various surgical methods were established during the twentieth century, which still are employed in the treatment of intractable Menière's disease, especially saccotomy and vestibular neurectomy but also intoxication of the labyrinth by intratympanic application of gentamicin. Despite the good results of such therapeutic regimens the basic pathological mechanism is still not fully understood. Since the description of an endolymphatic hydrops by Hallpike und Cairns in 1938 as a typical feature, there have been some observations of a possible infectious, allergic and autoimmunological (co)pathogenesis without enough proof to explain the disease in every case. This article aims to present the current scientific data, diagnostics and therapy of Menière's disease with special emphasis on surgical treatment options.

[Results in otosurgically treated patients with acoustic neuroma. Part 2: Hearing results after middle fossa approach]. / Ergebnisse otochirurgisch operierter Patienten mit Akustikusneurinom. Teil 2: Hörergebnisse nach transtemporaler Resektion.

BACKGROUND: The surgical therapy of acoustic neuromas has been seen critically lately. Alternative methods like the gamma-knife treatment open a wide field of discussion. PATIENTS: The results of 392 patients are presented operated on via a middle fossa approach between 1989 and 2004. RESULTS: Hearing preservation according to the 50:50 rule (hearing loss less than 50 dB in pure tone audiogram 500-1000-2000 Hz, speech discrimination>50%) was possible in 59.7% of patients. A hearing impression up to 90 dB was recognized in 83.1%, 16.9% of cases were deaf. There was no difference between tumors confined to the inner ear canal and tumors extending into the cerebello pontine angle. There was also no difference between tumors up to a volume of 100 mm3 and tumors between 100 mm3 and 500 mm3. CONCLUSION: Acoustic neuroma confined to the internal meatus or with little extend into the cerebello pontine angle (i. e. no contact to structures of the brain stem or vessels of the posterior fossa) are feasible for resection via the middle fossa approach. The possibility of hearing preservation and low morbidity makes it the treatment of choice for this group of tumors.

[Results in otosurgically treated patients with acoustic neuroma. Part 1: Facial nerve function after translabyrinthine and middle fossa resection]. / Ergebnisse otochirurgisch operierter Patienten mit Akustikusneurinom. Fazialisfunktion nach translabyrinthärer und transtemporaler resektion.

BACKGROUND: Regardless of all efforts to preserve a residual hearing, facial nerve function is still the primary issue for patients with acoustic neuromas. Since alternative methods like the gamma-knife treatment are of increasing importance, results after surgery have to be compared and discussed critically. PATIENTS: The results of 538 patients operated at the Dept. of Otolaryngology, Head and Neck Surgery, University of Wuerzburg between 1989 and 2004 are presented. 392 had surgery via a middle fossa, 146 via a translabyrinthine approach. RESULTS: Shortly after surgery 82.2 % of patients showed no facial palsy. 74 patients had an incomplete, 7 a complete paresis. After 12 months 144 of the patients operated on via middle fossa approach showed no palsy, 13 had an incomplete and one a complete paresis. Out of 76 patients operated on translabyrinthine approach 75 had no paresis, one had an incomplete, and no patient had a complete paresis. There was no difference between intrameatal tumors and tumors extending beyond the porus. Regular facial nerve function was seen in 93 %, postoperatively. Permanent paresis was seen in 0.4 % of cases. CONCLUSION: Acoustic neuroma confined to the internal meatus or with minor extension into the cerebello-pontine angle (i. e. no contact to structures of the brain stem or vessels of the posterior fossa) are feasible for resection via a middle fossa or translabyrinthine approach. The possibility of hearing preservation combined with low morbidity and good results of facial nerve function makes these approaches the treatment of choice for this group of tumors.

[Predictors of nodal metastasising in laryngeal squamous cell carcinomas as decision support for neck dissection: comprehensive analysis of literature]. / Prädiktoren der nodalen Metastasierung des Larynxkarzinoms als Entscheidungshilfe bei der Neck Dissection: Literaturanalyse zur Frage zuverlässiger Parameter.

BACKGROUND: A subset of advanced laryngeal squamous cell carcinomas (SCC) does not metastasize in regional lymph nodes (pN0). However, more than 30 % of tumors without signs of metastasizing in the clinical examination (cN0) show occult metastases. The guidelines of the German ENT-Society intend the extent of neck dissection (ND) depending on clinical stage of tumor and lymph nodes. If laryngeal surgery is followed by an adjuvant radiation/chemotherapy, ND is not always necessary. Histomorphological, immunohistochemical, or molecular parameters with predictive value for nodal metastasizing could support the planning for ND, especially in patients with cN0. METHODS: Within the last 20 years there were many publications concerning this problem. Herein, we analyzed the results of 455 publications. We have chosen studies regarding the predictive value of tumor stage, grading, peritumorous inflammation, invasion of lymphatic vessels, angioneogenesis, proliferation, overexpression of p53 or cyclin D1, inhibitors of cyclin-dependent kinases, growth factors, apoptosis, cell-adhesion, nm23, metalloproteinases, DNA/ploidy as well as tumor genetics. RESULTS: All examined parameters did not allow a fail-safe prediction of the risk for nodal metastasizing. CONCLUSIONS: Up to now, reliable predictors do not exist. The investigation of above mentioned parameters in pre-operative tumor biopsies is not helpful for the planning of ND in the stage cN0 (out of T1).

PKC isotype functions in T lymphocytes.

The main function of mature T cells is to recognize and respond to foreign antigens by a complex activation process involving differentiation of the resting cell to a proliferating lymphoblast actively secreting immunoregulatory lymphokines or displaying targeted cytotoxicity, ultimately leading to recruitment of other cell types and initiation of an effective immune response. In order to understand the physiology and pathophysiology of T lymphocytes, it is necessary to decode the biochemical processes that integrate signals from antigen, cytokine, integrin and death receptors. The principal upon which our work is based is to explore and identify gene products of distinct members of the AGC family of protein serine/threonine kinases as key players mediating cell growth regulation. Given the established important role of PKC theta as regulator of T cell fate and knowing that several other PKC isotypes are also expressed in T cells at a high level, we now summarize the physiological and non-redundant functions of PKC alpha, beta, delta, epsilon, zeta and theta isotypes in T cells. This review describes the current knowledge of the physiological and non-redundant functions of the PKC gene products in T cells.

[Malignant paranasal sinus tumors. Diagnosis, therapy and results]. / Maligne Nasennebenhöhlentumoren. Diagnostik, Therapie und Ergebnisse.

BACKGROUND: Malignant neoplasmas of the paranasal sinuses are rare and present usually in advanced tumor stage due to the lack of early clinical symptoms. PATIENTS AND METHODS: In the last 10 years, 63 patients with paranasal malignancies were treated at the Department of Otolaryngology, Head and Neck Surgery of the University of Würzburg. 33% of the patients showed an occupational exposition (wood-processing or metal industry). At the time of the first visit to our institution 95% of the patients presented with an extensive disease, staged T3 and T4. Adenocarcinoma (24%), squamous cell carcinoma (22%) and malignant melanoma (19%) were the most common histologies. Surgery combined with radiotherapy was the treatment strategy in 55 patients (87%). RESULTS: Patients with a complete surgical resection showed a higher 5-year-survival rate (77%) than patients with an incomplete resection (56%). In 38% (n=21) of the patients treated with surgery and radiotherapy, a local recurrence of the tumor was observed. This recurrence localised in the skull base and/or the orbita/periorbita occurred most frequently in the first (46%) or the second year (31%). CONCLUSION: The prognosis of malignant paranasal tumors depends mainly on the control of the local tumor growth. Modern strategies of surgical treatment in combination with radiotherapy need to be implemented in an effort to achieve a continuous remission.

[Injuries of the head and neck in suicidal intention]. / Verletzungen des Kopfes und des Halses in suizidaler Absicht.

BACKGROUND: Injuries of the head and neck with suicidal intention might create serious situations that require rapid and interdisciplinary treatment. METHODS AND PATIENTS: Twenty-seven patients with suicidal head and neck injuries were treated at the Department of oto-rhino-laryngology, head and neck surgery, University of Wurzburg/Germany, between 1991 and 2002. The medical histories were analyzed retrospectively. RESULTS: Twenty-three of them were male, 4 female. Mean age at time of attempted suicidal was 48 years (18 - 90). One patient was already treated for a psychiatric disorder. None of the patients had a suicidal attempt in the history. Nine patients suffered from a pharynx or larynx trauma after strangulation. Four out of seven patients with cutting or stabbing injuries showed a perforation of the upper airways. Nine patients had gunshot traumas, one of them with perforation of the pharynx. Thirteen patients underwent immediate endoscopy under general anaesthesia followed by a neck exploration in 3 patients. Six patients received a temporary tracheotomy. All patients were considered suicidal for the duration of stay in the ENT-department resulting in a permanent supervision. All patients were transferred to a psychiatric unit as soon as possible for further treatment. A statistically significant accumulation was observed during the last third of a year. Over the period of 11 years, suicidal injuries of the head and neck tend to occur more frequently. CONCLUSIONS: The presented study emphasizes the need of immediate surgical and intensive care treatment of patients with head and neck injuries due to suicide attempts as well as an adequate psychiatric supervision during as well as after the surgical treatment.

Direct interaction of the extracellular matrix protein DANCE with apolipoprotein(a) mediated by the kringle IV-type 2 domain.

Lipoprotein(a) [Lp(a)] consists of LDL and apolipoprotein(a), and has been shown to be a major, independent, risk factor for arteriosclerosis and thrombosis in humans. To further elucidate the (patho)physiological function(s) of Lp(a)/apo(a), we searched for new protein ligands, using the yeast two-hybrid system and employing the highly repetitive kringle IV type 2 (KIV-2) sequence from apo(a) as bait. The extracellular matrix protein DANCE [developmental arteries and neural crest epidermal growth factor (EGF)-like] recently implicated in atherogenesis was identified as an interactor. A direct physical interaction between DANCE and apo(a) was confirmed by co-purification of both recombinant proteins derived from culture supernatants of transiently transfected COS-1 cells. Furthermore, binding of human plasma-derived Lp(a) to recombinant DANCE was also observed. Finally, when monoclonal anti-apo(a) and polyclonal anti-DANCE antibodies were applied to tissue slices of atherosclerotic carotid artery, the two proteins were found to be co-localized in endothelial and smooth muscle cells, suggesting that they occur together in the arterial wall. However, as yet, the in vivo relevance and the possible functional role of this newly identified DANCE:Lp(a)/apo(a) interaction remains speculative.